We narrowed to 1,052 results for: FER

... dates. Buffers Buffers are a concern for conjugating and signaling molecules, as some buffer components...heavy chain constant region” or Fc. Different isotypes play different roles in the immune response, and ...removed from the buffer before the reaction. However, it can be added back to buffers after the conjugation...of colors activated by different wavelengths, allowing for easy differentiation between multiple antibodies... In some cases, it may be preferable to buy small aliquots of many different antibodies and do batch validation...an antigen, while the specific binding site is referred to as an epitope. Production Antibodies are typically...typically produced in one of three different methods: in animals, in cultured immune cells, or through...

... via a different plasmid. In order to produce rAAV vectors, three plasmids are needed: Transfer plasmid...s Molecular Biology Reference . Production The three production plasmids (transfer, packaging, and helper...2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central...immune response. These features have made AAVs the preferred choice to be used as viral vectors for gene therapy...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites or...recombination of unstable regions, like the ITRs found in transfer plasmids. This will ensure that the repeats will...

...three different plasmids containing the transfer, envelope, or packaging components. The transfer plasmid...packaging genes. However, they are different viruses and require different isoforms of these packaging components...type. From an experimental standpoint, the main difference between lentiviruses and gamma-retroviruses is...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA ... gene, due to a wide range of infectivity for different cell types (known as tropism). Figure 2: Gamma-retroviral...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites or...

...There are many different lysis buffers available commercially, and most common buffers have recipes available...Types of transfers Your transfer options are wet, semi-dry, or dry/electroblot. A wet transfer is the most...  Note that each type of transfer requires different equipment. Wet transfers can be done in an insert... while semi-dry transfers require a semi-dry transfer cell. Dry/electroblot transfers only work in the...is typically sufficient.  Lysis buffer When choosing your lysis buffer, consider the subcellular location...and Tris gels run proteins in different patterns, due to their differing chemistries. You therefore cannot... 12% 10–250 kDa Different sizes 4–12% gradient 4–250 kDa Different sizes 4–20% gradient...

...many different Cas proteins available, and researchers can adapt these experiments for different organisms...Desired Genetic Manipulation Different genetic manipulations require different CRISPR components. Selecting...editor variants often have different optimal editing windows. Be sure to refer to the literature associated...Activation/interference Needs good primer design Western blot Knockouts Activation/interference Needs a ...Expression and Delivery Validate Genetic Edit References CRISPR ( C lustered R egularly I nterspaced S...have expanded the CRISPR toolbox to include many different types of edits, including point mutations, base...Base editor (dCas9 or Cas9 nickase fusions with different base editing proteins) Many base editors fused...

... (A to G). Different serotypes exhibit varying tissue tropisms, meaning they preferentially infect certain...from different species. Used to alter infectivity and tropism. pShuttle Class of shuttle/transfer vectors.... These genes are now supplied in trans , via different plasmids or the packaging cell line, which not...need two plasmids: Shuttle plasmid (also known as transfer plasmid) — containing the transgene of interest...recombination between the transfected adenoviral transfer plasmid and the E1 region integrated in the packaging...vectors Third-generation rAdV vectors are also referred to as "gutless" or high capacity AdV (HCAdV) vectors...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...

...melting profiles (Dwight, 2011).  Reference Genes  Choosing a set of reference genes deserves more attention...attention than it usually receives. Different reference genes have shown to have as much as sixfold variation...Passive Reference Dyes Whatever your pipetting method, some variation will occur. A passive reference dye,...Liu and Saint No Raw Data Reference and target genes can have different efficacies DART-PCR No ...will soon become a qPCR master. References and Resources  References Biassoni R; Raso A. (eds.), Quantitative...mRNA, which can be used as a proxy measure for differences in protein expression. (If you're just getting...assume that qPCR can accurately detect expression differences of twofold or greater with good statistical significance...

... wet transfer system the sandwich is completely submerged in a tank filled with a transfer buffer formulated...systems efficiently transfer a range of protein sizes in minutes without any transfer buffer. These systems...electric current. The wet transfer process is very effective in transferring a broad range of protein ...overnight.  In contrast, a semi-dry transfer only requires enough buffer to saturate the sandwich and typically...validated for immunoblotting. Different immunoassays detect proteins in different states. For example, in immunoprecipitation... other antibody topics. Resources and references References Aydin S (2015) A short history, principles...western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using antibodies...

...heterogeneous population, cells behaving differently (e.g. different growth properties, migration abilities... modified cells are transferred into an animal and the behavior of the transferred cells is followed. ...targeting all differentially expressed genes that are known drug targets, or all differentially expressed ...best gRNAs by the depositing lab. The different labs use different algorithms to calculate which would ...were not found in the reference library. It’s not uncommon that genes have different names, and the software...generation sequencing to identify which gRNAs are differentially expressed in the separate groups and, as a ...is not targeted by only one gRNA, instead many different gRNAs are used for each gene. At the moment, 13...

...Addgene blog Plasmid Preps: Different Purity, Different Quantities, Different Uses Plasmids 101: Optimizing...lysis buffer ratio and result in incomplete lysis. You may be able to scale the amount of buffers used ...TakaraBio) offer yeast-plasmid specific kits.  Medium/High-throughput – Many manufacturers offer bead- and...quantity.   Figure 1: Plasmid preps fall into different categories based on their relative qualities and...fairly self explanatory and achieved by performing different sized preps, such as miniprep, midiprep, etc. ...what do we mean by DNA quality? There are a few different issues that can arise during the plasmid purification...which can happen if you leave the sample in a lysis buffer for too long or if you get a little overzealous...

...stable expression of the transgene   Transfer plasmids The transfer plasmid is the vector you clone your...the control of two different promoters. Both the single and dual promoter transfer vectors can be used...genetically modify primary cells. Amongst the different vectors used, lentiviral vector constructs have...background Until the development of viral vectors, gene transfer into mammalian cells often relied on chemical ...These techniques are still used and efficiently transfer plasmid DNA efficiently into dividing immortalized...lines but reach their limits when it comes to transferring DNA into primary cells or non-dividing cells... particle (Gag, Pol, Tat, Rev and Env) and the transfer plasmid encoding the insert of interest as well...

.... While these differences may seem subtle they result in a key physiological difference between lenti-...3rd-generation transfer vectors apart? A3: Addgene defines 2nd- or 3rd-generation transfer vectors based...packaging systems differ between 2nd- and 3rd-generation vectors? What does this mean for transfer vectors? A4...protein and consequently are considered safer. Second-generation transfer plasmids MUST be packaged with a 2nd-generation... Are transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer plasmids...which is rarely more than 30% of the transfer plasmid. Many transfer vectors are SIN. Packaging plasmid...ingredients: the lentiviral packaging vector, a transfer vector including the gene of interest, and the...

...Alternatives Resources Plan Your Experiment Web References PAM Sequences Glossary Publications CRISPR Overview...techniques . Collectively, these techniques are referred to as CRISPR ( C lustered R egularly I nterspaced...measure of fidelity, so sometimes they will be referred to as increased fidelity enzymes. hfCas9s were...pyrimidine-to-pyrimidine) Base transversions — edit to different classification of nucleotide base (purine-to-pyrimidine...opposite strands Prime editing and base editing both offer high efficiency and low indel formation compared...genetic changes, including moving entire genes to different locations, by combining other gene editing mechanisms...DNA-targeting of CRISPR-Cas systems. You may also see them referred to as INTEGRATE ( I nsertion of T ransposable ...

...Mobile-CRISPRi: a modular approach for CRISPRi transfer Mobile-CRISPRi offers two advantages over existing bacterial...chromosomal copy of the RP4 transfer machinery mobilize these plasmids for transfer to recipient cells during...disease. However, the rate of transfer was variable. They measured transfer rates by comparing the number... P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates are better suited for...Mobile-CRISPRi plasmids at Addgene! References Auchtung, Jennifer M., et al. "Biology of ICEBs1, an integrative...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria...diverse bacteria species. Existing protocols can transfer CRISPRi into a single bacterial strain, such as...

...approach is RNA interference (RNAi). The most common method is transfection of short interfering RNA (siRNA...For CD365, they examined two different antibody clones produced by different vendors. The specific epitope... panels, where different manufacturers may have the same clone available with different conjugates. Summary...commercial antibody manufacturers. References and resources References Ayoubi, R., Ryan, J., Biddle, M. ...include a discussion of the Human Leucocyte Differentiation Antigens (HCDM) workshop approach, which focuses...one size fits all. The approaches used will be different depending on the sample and target type. To start... For intracellular targets, we usually test 3 different fix/perms (methanol, PFA-saponin, PFA-triton)....

...light-emitting luciferase and a light-sensing optogenetic element. When the luciferase substrate (luciferin) is ...efficiencies. Depending on the type of luciferase used, the luciferin substrate may be the native coelenterazine...activity in neurons LMO Luciferase Opsin Ligand Effect Outcome (in neurons) Reference LMO3 sbGLuc VChR1 CTZ...RASSLs DREADDs PSAMs LMOs Plan Your Experiment References Chemogenetic tools are actuators for specific... of chemogenetic receptors. There are several different types of DREADDs that can be broadly classified...inhibitory KORD DREADD was developed to respond to a different ligand, Salvinorin B (SALB). Scientists have shown...Receptor Effector Ligand Effect Outcome (in neurons) Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO...

..., A mutant Renilla luciferase, catalyzes the oxidation of its substrate, luciferin, which releases energy... neomycin or luciferase) to create a final lentiviral vector containing up to 5 different elements. Lentiviral...bioluminescence resonance energy transfer (BRET), whereby photons emitted by a Renilla luciferase variant are used to...alternative to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids... issues with fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash...pHlash, that use Bioluminescence Resonance Energy Transfer (BRET) were developed by the Johnson Lab . Characteristics...energy as a photon. The photon energy is transferred to the acceptor fluorophore, a circularly permuted...

... directed differentiation To understand how each human TF can contribute to differentiation, the Zhang...atlas. Computational analysis was used to infer differentiation trajectories and identify both up- and down-regulated... scientist at [email protected]! References and Resources  References Joung J. et al., A transcription...patterns determine everything from stem cell differentiation fate to tumor suppression (and most everything...construct. Cells were cultured following a differentiation protocol before performing single-cell RNA ...embryonic stem cells. Transduced stem cells were differentiated and single cell RNA-seq was performed to map...    This atlas linked TF expression to differentiation outcomes as well as global transcriptional ...

...most organisms on Earth, but different organisms have different preferences for the codons they use to ... organisms have different codon usage biases? The reasons for varied codon preferences among organisms...poorly charged tRNAs, different regions of the protein may be translated at different rates. The ribosome...Organisms often need to express genes at different levels under different conditions. With varied codon usage...constraints have molded which codons are used preferentially in which organisms - organisms have codon usage...your organism of choice. Unfortunately, codon preferences make it so you cannot choose among the possible... evolutionary constraints that lead to these preferences and what can we do about them? Read on to find...

...your TTO have a question? Refer them to Addgene’s Tech Transfer Team: techtransfer@addgene.org Congratulations...Addgene Help Center: Tech Transfer Technology Transfer Information Tech Transfer News: A Reintroduction ...Addgene account for Tech Transfer Offices. They’re packed with tech transfer resources (summaries, electronic...stats and strategies with tech transfer offices. Drop us a line at techtransfer@addgene.org. Albert Einstein...requests from Addgene MTA: material transfer agreement Technology Transfer: the exchange of intellectual property...addgene.org identifies the hold-up. Your Material Transfer Agreement is stuck in “pending.” Fear not, there... to make MTA turnaround faster with technology transfer offices (TTOs). The latest of these efforts is...

...utilized mice to study how different mutations in leukemia impact different treatment regimens (Zuber ...example, the Stainier lab studied the proliferation and differentiation of β-cells in the the developing zebrafish...β-cells that originate in different regions of the pancreas and produce different levels of insulin (Hesselson...mouse (Mus musculus) Let’s start with the most preferred mammalian model organism of many researchers: ...injecting mice with retroviral vectors that harbored different mutations found in leukemia, the Lowe lab was ... gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces cerevisiae...at a time. Zebrafish embryos are also laid and fertilized externally, allowing scientists to easily manipulate...

...test assesses the sum of differences between pairs). Samples arising from different subjects or populations...test assesses the difference of the group means). Unpaired t-tests can also be referred to as two-sample...significant differences between the means of your groups. However, it will only tell you if differences exist...samples are different from each other, or if every permutation of pairs within the five is different. To identify...groups to determine if they are statistically different. They are classic parametric tests and one of ... whether or not two samples are statistically different than one another, a two-tailed test should be ...control group. If you only care if one group is different in a certain direction (i.e., you just want to...

... Opsins Optical Switches Plan Your Experiment References The field of optogenetics integrates optics and...naturally occurring microbial opsins that respond to different wavelengths of light, like blue or yellow light...light. These various opsins also initiate different electrochemical responses, such as nonspecific cation... blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance ...optimization, and chimeric fusion of domains from two different ChRs. These ChR variants still function as light-gated...Color of activation light There are a variety of different activation wavelengths, ranging from blue to yellow...rendering the experimental procedure less invasive. Different activation wavelengths also make it possible to...

..., Addgene uses 'labeling' to refer to antibodies and 'staining' to refer to small molecule dyes. Indirect...first fixed and then permeabilized, using two different buffer solutions. Fixing causes protein cross-linking...nucleus, a different fix/perm kit is required (to be precise, the fixing solution is different while the...plot (Figure 4b). Through this method, differences in proliferation between cell populations can be visualized...divisions).   Figure 4: Proliferation dyes can visualize proliferative capacity and speed of cells...lineage markers, like many of the cluster of differentiation (CD) proteins found on immune cells. However...use 'labeling' and 'staining' interchangeably to refer to any technique that combines a marker with a signal...

...maintaining the undifferentiated “stem” state and, Their potency or capability to differentiate into specialized...special types of cells that can develop or “differentiate” into more specialized cells with specific functions...function adequately [1]. Stem cells’ ability to differentiate into multiple cell types makes them useful models... cell has the ability to produce all of the differentiated cells in an organism, including the embryonic...Pluripotency: When a cell has the ability to differentiate into cells of any of the three germ layers: ...detail below [2]. Multipotency: When a cell can differentiate into multiple cell types of the same lineage...name suggests, cells under this category can differentiate into only one cell type. Muscle satellite cells...

... virus can be stored at 4 °C and buffer exchanged the next day. Refer to the AAV Iodixanol Gradient Ultracentrifugation... AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid...help mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest ...more details. Pro tips I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple...to grow multiple bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen...restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, like NEB Stable...

...created phages. Transfer of this DNA to another cell is referred to as transduction. Transferred DNA once inside...donor can transfer genetic material to the recipient bacterium. The genetic material transferred is commonly...Horizontal gene transfer (HGT) is the movement of genetic material between organisms. It plays a key ...unique to the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...Various factors promote natural transformation in different bacteria such as growth phase of the cells (Baltrus...plasmid, the genes on the plasmid can give them different phenotypes. For example, this plasmid encodes ...cell. Once inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages ...

...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...engineered γ-retroviruses differ from lentiviruses? There are a few important differences between engineered ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...Vectors 101 eBook, 1st edition.  References and resources References Swift S, Lorens J, Achacoso P, Nolan...following components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA ...sequences, which facilitate integration of the transfer plasmid sequences into the host genome. Typically... the host genome upon viral transduction. Most transfer plasmids available from Addgene were derived from...

...Tris-EDTA (TE), just Tris buffer, or some other variations. Does it make a difference? Short answer: yes. This...choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose could...TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving the stability of your preparation...can be Tris buffer (with no EDTA) or water. These are two of the choices that companies offer for the elution...preparation can be eluted in water or in your buffer of preference, and it will be stable for years. Plasmid... preparation for a long time. Tris buffer controls the pH, while the EDTA chelates any divalent cations...through their kit. The advantage of using a Tris buffer is that it allows better control of the pH preparation...

...through horizontal (or “lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation...conjugation (direct transfer from one cell to another). Bacteria need special fertility genes to be capable...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...Here at Addgene, we often refer to plasmids as lab or experimental tools. They certainly are very handy... evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001) Beneficial...humans only noticed last century! Nevertheless, the transfer of plasmids carrying these genes between bacteria...Historically, colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.” Virulence...

...host range is determined by the different hosts that a plasmid can transfer to, replicate in, and persist...stable, can be easily cloned in different bacteria, and can be transferred to various experimental hosts... that can replicate in different bacteria and express proteins within different host organisms. pBBR1-...recombinant vectors at Addgene!   References and resources References Doran, K. S., Konieczny, I., & Helinski...host range, identify a few of the features that confer broad host range in plasmids in nature, and provide... they carry among organisms belonging to many different phylogenetic groups (Jain and Srivastava 2013)...be used by scientists across disciplines.  What confers broad host range in plasmids? Not all plasmids ...

...your poster Check conference guidelines To begin, you should always follow conference rules when designing...useful to use two different font styles; one for your section titles, and a different one for the body ...distinguished. The UCLA Library offers some Powerpoint based templates with different column numbers for a good... the most of your conference attendance Read all of our blog posts about conferences .... But, before you can show off your work at a conference, you need to first make your poster. While a ...understand that presenting your work in poster format differs in many ways from writing a manuscript. If you ...you have ever attended a scientific conference, or even a lab recruiting session at your university, you...

...not transfer precisely evenly across the entire blot. So how can you accurately compare different lanes...assuming that all differences in the loading control are due to loading and transfer errors — not to any...reflect massive differences in protein abundance. Validating the assumptions for different loading controls...detection range be linear! Resources and References References Aldridge, G. M., Podrebarac, D. M., Greenough...errors like uneven pipetting or irregular membrane transfer. Figure 1: A loading control is a type ...Researchers use later steps in this chain (blue) to infer or calculate things about earlier ones. In practice...relationship poses problems for proteins of very different abundance, consider the following example gel....

...There are many different flavors of MTAs, but the Uniform Biological Material Transfer Agreement (UBMTA...process that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer office is...for more information. Happy transferring!    Many thanks to our Tech Transfer team members Maura Coughlin...Addgene, then you’ve been involved in the material transfer agreement (MTA) process. Even though this process...curious about the hows and whys of the material transfer agreement, read on!  What is the MTA?  An MTA...UBMTA) is one of the most commonly used ones for transfers between academic institutions.  What is the UBMTA...the creation of a standardized agreement on the transfer of biological materials in 1995: the UBMTA. Because...

... deposited by Promega here.    References and resources   References Allikalt, A., Purkayastha, N., Flad... scientists. Seeing inside these specimens has offered illumination on biochemical processes that are ...Figure created with BioRender.com. Fluorescence refers to the emission of light by a substance (fluorophore...energy due to non-radiative energy losses. This difference in emission results in a slightly longer wavelength...phenomenon is known as the Stokes Shift, or the difference in wavelength between peak excitation spectrum...vary in their chemical structure, leading to differences in their excitation and emission wavelengths,...target specific organelles like mitochondria, and different dyes can result in varying cytoplasmic background...

...Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers...bioluminescence resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite...problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently...Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent technology...need for an excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in ...technique’s versatility and adaptability to many different contexts. Improving Nano-lanterns In 2015, Takai...eNL constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition...

...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which... acceptor fluorophore. The difference for BRET is that the energy transfer is between a luminescent donor...number of different experimental approaches. They first assessed the activation of different members of...response to different agonists. These studies showed the promiscuity of GPCRs for different G protein transducers...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002...a vast number of signaling cascades, inducing different cellular responses. One of the biggest advantages...measure the physical association of combinations of different Gα, Gβ, and Gγ subunits. When inactive, these ...

...control many cellular processes such as differentiation, proliferation, and apoptosis. In humans, mutations...Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly luciferase. The bait and prey...variable availability of enhancer sequences in different chromatin states. Alexander Stark and colleagues... were deep sequenced to identify the gRNAs that confer protection to WNV. From this screen the authors...process of cloning, combining, and testing these different parts can be quite arduous. The cloning process...consists of a variety of plasmids containing many different promoters, RBS’, coding sequences (CDS), and terminators...To facilitate innovative cancer research, the Reference Reagents Group of the NCI RAS Initiative, led ...

... variables.    Serotypes Serotype refers to the different protein compositions of AAV capsids, which... a virus enters the cell. Different serotypes utilize a number of different receptors, co-receptors, and...burdensome in the brain where many different cell types, or even different phenotypes of the same cell type...working in several different systems or want to transduce a large number of different cells. Promoters ...doi.org/10.3389/fnmol.2014.00076 References and Resources References Aschauer, D. F., Kreuz, S., & Rumpel...validation and optimization. Although there are different ways to ultimately achieve success, we find it...validation first.  AAV-mediated gene expression can differ extensively depending on the experimental system...

... However, the two different approaches allow for optimized strategies for differing amounts of target ...urea-containing lysis buffer. Alternatively, the protein can also be eluted using a low pH buffer such as 0.1M ...for other antibody topics.     References and resources References:  Björck L, Kronvall G (1984) Purification... appropriate for most situations.  A slightly different method is desirable if the antibody is expected...Immobilization is the key to immunoprecipitation and simply refers to the process of anchoring an antibody, often ...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ...than those of agarose beads, despite the size difference. Magnetic beads are particularly advantageous...

...target primaries produced in different species and to label targets with different colors. Multiplex immunofluorescence...Speaking of different colors, having a palette of secondary antibodies in a few different species and ...dimensional epitopes. Different sample processing steps can alter epitopes in different ways, so antibodies... a successful IF experiment.    References and resources References Im K, Mareninov S, Diaz MFP, Yong...immunohistochemistry (IHC). All three of these terms refer to the use of antibodies to visualize a target within...essentially just more general terms that can also refer to assays using non-fluorescent visualization methods...indirect immunofluorescence There are a couple different approaches to be familiar with when talking about...

... IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation and had ...number, generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent... brain.  Figure 1:Schematic diagram of different FingR constructs and experimental results. Image... of the cell. By combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory... proteins system for Caenorhabditis elegans By Jennifer Tsang Fluorescent proteins (FPs) traditionally...The team validated this tool by targeting six different genes whose protein products have distinct localizations...Figure 2: Split-wrmScarlet labeling proteins with different subcellular locations. Image from Goudeau et al...

...blot Run your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize...before rinsing with wash buffer, as dry PVDF won’t evenly re-absorb aqueous buffer. Be sure to check the ...from having to run a whole new gel and membrane transfer. Let’s get started! Before you begin In the best...abundant. If the abundance of your targets isn’t too different, consider antibody affinity: stronger binding ...should not compare the strength of bands from different rounds of detection. Finally, stripping and reprobing.... Unlike PVDF, they can be returned directly to buffer without a re-wetting step. Important: do not dry...so you’ve dried your membrane and returned it to buffer, you’ve run and imaged your western blot with your...

...worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities depending... Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...make the virus. Viruses can be produced in many different cell types, spanning both prokaryotic and eukaryotic...However, some viruses can be produced in multiple different cell types, including plants, mammals, and insects...improve your current methods, understanding the differences can help you make an informed decision. Mammalian...AAV and lentivirus requires three plasmids — a transfer plasmid containing your gene of interest plus ...standard to do only one passage (transfection). This differs from typical insect cell preparations, which we...

...required, through mitochondrial transfer. In the two decades since this transfer was first proposed, researchers... astrocytes Nerve-to-cancer cell mitochondrial transfer tracker Red calcium indicators PinkyCaMP and FRCaMPi...better or for worse. To better understand this transfer, the Simon Grelet Lab developed MitoTRACER, a ... cancer and melanoma models, showing that this transfer increases the metabolic and metastatic capacity...it is a diverse tool that can be used in many different systems, both in vitro and in vivo. Find MitoTRACER...2023). The power and potential of mitochondria transfer. Nature, 623(7986), 283–291. https://doi.org/10.1038...-z Hoover, G., et al. (2025). Nerve-to-cancer transfer of mitochondria during cancer metastasis. Nature...

...expression. Gene isoforms refer to mRNAs generated from the same gene but with different untranslated or coding...activated cell sorting (FACS) to isolate differentiated from undifferentiated cells at the end of the screening... to study neuronal differentiation, it could also be applied to the differentiation of other CNS cell ..., we’ll highlight these recent advancements and offer commentary on their application to neuroscience ...straightforward to use CRISPRa to selectively target these different transcriptional start sites in order to better... expression of Bdnf resulted in hundreds of differentially expressed genes between Bdnf I-overexpressing...which transcription factors promote neuronal differentiation (Liu et al., 2018). The pooled nature of this...

... immune system make different isotypes of antibodies that recognize many different epitopes of the target... and protein G each have different affinities for IgG subtypes from different animals so it’s important...successful next experiment!     References and Resources References Ascoli CA, Aggeler B (2018) Overlooked... cover polyclonal antibodies, one of the many different types of antibodies available (others you’ll encounter...polyclonal antibodies from the serum There are a few different ways the antibodies can be isolated from the serum...by an animal are specific for the immunogen. A different type of purification can be performed to isolate...experiments where antibodies raised in several different species are present. One common example is using...

...Castle et al., 2014). Differences between AAV serotypes may be also linked to differences in viral uptake by...these different AAV vectors into a brain region, the AAV particles will infect neurons at different ratios...architecture, in particular the connections between different brain regions. For example, communication between...2015). Directional flow of information between different parts of the brain is mediated via individual ... for degradation. These two pathways also use different cytoskeletal machineries to facilitate transport...while anterograde transport relies on kinesin. Different rates of speed for various forms of retrograde... or have been genetically engineered to have differences in their capsid proteins. For a brief overview...

...Your AAVancement could be next!   References and Resources References Braun, C. J., Adames, A. C., Saur...allows for a wide variety of screens, including interference, activation, and knockout. Most CRISPR screens...CRISPR screens can be carried out using a few different methods — in vivo, in vitro, and ex vivo. The .... The go-to in vivo option AAV has become the preferred delivery method for in vivo CRISPR screens. AAV...titers equal to those achieved with lentivirus but offer an improved safety profile, customization, and enhanced...mouse tissues. Safety AAV and lentivirus have different safety profiles that often affect which is chosen... cause bad reactions in mice. Customization AAV offers a level of customization that is unmatched by other...

...antibodies are all of different species/isotypes and that each secondary has a different color fluorophore...a comment below. Happy imaging! References and Resources References Andrews, N. P., Boeckman, J. X., ...be familiar with immunofluorescence (IF, often referred to as immunocytochemistry (ICC) when the sample...mixture of secondary antibodies, and image all the different colors in one session – fast and easy! Just make...Trimmer Lab, using recombinant antibodies with different isotypes. (A) Neocortex labeled with anti-pan-...make sure you are using primary antibodies from different species or isotypes, so that the secondary antibodies...to align the images of each labeled target as different color channels and analyze the results. Depending...