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We narrowed to 986 results for: igh

Showing: 101 - 150 of 986 results

  1. All Antibodies

    Type
    Collection
    ...for Protein Innovation Collection : Antibodies to highly conserved proteins and other difficult-to-target...

  2. CRISPR Plasmids - gRNAs

    Type
    Collection
    ... target various genes or genomic regions. It is highly recommended that you determine if your cell type...

  4. TALEN Engineering

    Type
    Collection
    ...FLASH (Fast Ligation-based Automatable Solid-phase High-throughput) assembly method (Reyon and Tsai et al...

  5. CRISPR Plasmids - Base Edit

    Type
    Collection
    ... available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have been ...

  7. Microbiology Resources

    Type
    Collection
    ... in bacterial genome editing is not always straightforward. Read our CRISPR Methods for Bacterial Genome...

  9. p53 Pathway

    Type
    Collection
    ... The p53 name arose from its observed molecular weight (53 kDa) when run on an SDS-PAGE gel. Cancer Impact...

  10. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...and Cpf1, which require that each DNA target have high sequence identity to the spacer sequence and contain...

  11. Arf GTPase Family

    Type
    Collection
    ... Casanova, J. E., Cherfils, J., Dacks, J. B., Lambright, D. G., Lee, F. S., Randazzo, P. A., Santy, L....

  12. Control AAV Preps

    Type
    Collection
    ...We provide high quality AAV preps from select plasmids in the repository. Browse our control AAV collection...

  13. Ras Pathway

    Type
    Collection
    ...erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 PAK PAK1 ...

  15. Lentivirus Plasmids

    Type
    Collection
    ...transgene expression, contains intron that gives you high expression. Trono 12254 pWPI 2nd EF-1alpha driven...

  16. CRISPR Plasmids - Tagging

    Type
    Collection
    ...vector serves as a backbone to clone the left and right homology arms. More details on how to use this plasmid...

  18. Retrograde AAV viral preps

    Type
    Collection
    ...Looger 106174 pAAV.hSynapsin.SF-iGluSnFR.A184S Syn Tight affinity glutamate sensor Glutamate sensor Looger...

  19. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...Protein Activation Library 113345 Activation Human Wright 3rd 7-14 58,570 Human miRNA CRISPR Knockout Library...

  20. Plasmids for Stem Cell Research

    Type
    Collection
    ... Lin28A to somatic cells from separate plasmids Highly Efficient Reprogramming to Pluripotency and Directed...

  21. Weighing Reagents Protocol

    Type
    Protocol
    ...reagent you will be weighing out, a weighing boat or weighing paper to contain the reagent while on the...that you are weighing by looking for a weight range on the scale. Make sure that the weight of the material...out the material you are weighing onto the weighing boat or paper. If you weighed out too much material,... up. Discard the weighing boat or weighing paper in the trash if the material weighed is not biohazardous... Protocols Weighing Reagents Weighing Reagents Protocol Intro ... to handling the materials. Equipment Weighing boats or weighing paper Balance Scoopula or spatula Container...button. Before weighing out your reagents, determine the amount that you need to weigh. Gather the reagent...

  22. Using a Light Microscope Protocol

    Type
    Protocol
    ...The route the light follows from the source to your eyes is called the light path . Light travels from ... Protocols Using a Light Microscope Using a Light Microscope Intro ... to encounter: the compound light microscope. As the name suggests, light microscopes take advantage of...compound light microscope with the main components labeled: Figure 1: Diagram of a compound light microscope...from the light source through a condenser that helps concentrate the light onto the sample, and then through...objective! Once on the higher objective, you will likely need to re-adjust lighting and focus. Increase ...basics of how to use a light microscope. Protocols...

  23. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing)... under either no excitation light, UV light or LED blue light.  Highlights: This is a one-pot, one-temperature...heat block, LED blue light illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK...captured by an antibody further along on the strip. Highlights: Of the 83 known COVID-positive samples tested... extraction but that may work only if there is a high enough (105 copies per microliter) viral load in...lateral-flow readout slip that allows rapid detection.  Highlights: Cas13a is not catalytically active in the presence... duplex and will not give a positive result for highly similar viruses. Unlike Cas12a, Cas13a does not...

  24. Optogenetics Guide

    Type
    Guide
    ...Microbial Opsins Opsins are light-gated ion channels or pumps that absorb light at specific wavelengths. ...respond to different wavelengths of light, like blue or yellow light. These various opsins also initiate...also a blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance...Channelrhodopsin from Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum... 566 ArchT Light-activated outward proton pump from Halorubrum strain TP009. Improved light sensitivity...binding domain, allowing light to activate transcription. Phytochromes and light oxygen voltage (LOV) domains...) BLUF domains bPAC Light-activated adenylyl cyclase from Beggiatoa sp. for light-induced cAMP modulation...

  25. Antibody Guide

    Type
    Guide
    ...the Fab fragments contain highly variable regions, shown in light blue and light orange, that bind to a ...are light-sensitive molecules which emit a specific wavelength of light after excitation by light of a...pair the brightness of the fluorophore with the abundance of the antibody target. If a bright fluorophore...molecular tools used in the lab to specifically and tightly bind to other proteins. Antibodies, produced as...-efficient way to produce antibodies, with the highest variability in antibodies produced. Cells - Antibodies...protein chains: two large heavy chains and two small light chains. These four domains come together to form... to be produced in a single individual, giving a high probability that an antibody will be able to recognize...

  26. CRISPR Guide

    Type
    Guide
    ...joining (NHEJ) pathway and the less efficient but high-fidelity homology directed repair (HDR) pathway....also enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no ..., which are about the same size as SaCas9, with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum... downstream of the target site (termed left and right homology arms). The length of each homology arm ...available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...strands Prime editing and base editing both offer high efficiency and low indel formation compared to HDR...favor insertion in the T-RL direction, with the right homology end closest to the target site. Type V-...

  27. Adenovirus Guide

    Type
    Guide
    ...simplicity of production, and higher titers. However, the risk of RCA formation is higher for first-generation ...rAdV vectors are also referred to as "gutless" or high capacity AdV (HCAdV) vectors. Here, all viral sequences...adenoviral backbone plasmid have matching left and right homology arms which facilitate homologous recombination...the shuttle plasmid, but this method results in a higher background of non-recombinant adenoviral plasmids... used for rAdV vector construction, due to their high gene transfer efficiency. Other serotypes like Ad26...immunogenicity. For example, Ad5/35 and Ad5/26 combine the high transduction efficiency of the Ad5 serotype with... particle. Vaccine Development rAdV vectors are highly immunogenic because they activate both innate and...

  28. Plan Your Experiment

    Type
    Guide
    ...for a given experiment. In the table below, we highlight the most common categories of genetic manipulations...joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity Dual-nickase...editors Editing efficiency can be low, but generally higher than HDR pegRNA optimization can be challenging...location. Positioning your chosen Cas enzyme at the right spot within the genome relies on proper gRNA design...design a donor DNA to have the appropriate left and right homology arms. See Addgene's HDR blog post for more...guides that target closer to the edit site yield higher editing efficiencies. The template sequences can...specificity, consider using an engineered Cas enzyme with higher fidelity. In addition to off-target activity, it...

  29. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...episome which can concatemerize, producing high molecular weight structures that are maintained extrachromosomally...hybrid capsid derived from eight different serotypes. AAV-DJ displays a higher transduction efficiency in...on transgene expression and the expression of neighboring genes. rAAV episomes can develop chromatin-like...in vitro than any wild-type serotype, and a very high infectivity across a broad range of cell types in...Brain Connectivity Atlas and the Janelia Campus MouseLight project . rAAV vectors have also been developed...signals of cells (commonly neurons) in response to light. The short coding sequences of channelrhodopsins...wild-type). Another split AAV method which has shown higher efficiency in delivering large genes makes use ...

  30. Chemogenetics Guide

    Type
    Guide
    ...opto-chemogenetic fusion proteins of a light-emitting luciferase and a light-sensing optogenetic element. When...neurotransmission. This, and the fact that GPCR ligands can have high potency, which is potentially important for dosing...off-target effects in vivo and the receptors exhibited high levels of constitutive activity, making them less...luciferin) is added, the luciferase enzyme generates light that activates the opsin. The opsin may excite or...illumination), making LMOs a versatile option. Coupling brighter bioluminescent modules, such as luciferase-fluorescent...example, a study in rats using an hM4Di receptor might consider an alternative DREADD ligand such as DCZ... chemogenetics by using physical and biological light sources for opsin activation. Proc Natl Acad Sci...

  31. Molecular Biology Reference

    Type
    Guide
    ...insert expression (i.e., a strong promoter directs high expression, whereas weaker promoters can direct ...determines the type of plasmid and dictates how it might be used in the lab. Below are some common plasmid...particles, that can infect your target cells at a high efficiency. Addgene's expanding viral service offers...Next Generation Sequencing (NGS) comes in. NGS is a high-throughput, multi parallel sequencing platform that... Plasmids 101: 5 factors to help you choose the right cloning method Plasmids 101: Restriction Cloning...

  32. Cloning

    Type
    Guide
    ...choosing the right cloning method for your research project . The following guide will highlight several of...

  33. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...for fast and efficient generation of large-scale, high-titer preps. This is thanks to established retroviral... on a case-by-case basis. Retroviruses carry a slightly elevated risk of integrating into proto-oncogenes...clinical-grade gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing...

  34. Science Guides

    Type
    Guide
    ...immune system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based...technologies developed for optogenetics research utilize light to detect, measure, and control molecular signals...

  35. Lentiviral Vector Guide

    Type
    Guide
    ...J., Bae, S., & Hur, J. K. (2021). CReVIS-Seq: A highly accurate and multiplexable method for genome-wide...Kochenderfer, J. N., Norberg, S. M., Hinrichs, C., Highfill, S. L., Somerville, R. P., Panch, S. R., Jin, ...

  36. Guide to Using Pooled Libraries

    Type
    Guide
    ...additional hypothesis-directed experiments. As you might have guessed from their name, pooled libraries are...

  37. Promoters

    Type
    Guide
    ...polymerase) to bind to a promoter. Transcription is tightly controlled in both bacteria and eukaryotes. Promoters...

  38. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ...limit of a light microscope. Due to the wave nature of light, an infinitely small point of light will blur... to provide high 3D spatial resolution while also avoiding the toxic effects of high light doses to the...fluorescence microscopy provides a high contrast image in which a bright signal overlays a perfectly black...placed to only allow light from the focal plane to reach the detector and all light from above or below...for example, red light instead of blue to excite a molecule of GFP. Because red light has approximately...approximately half the energy of blue light, two photons of red light are needed to excite GFP compared to only ...objective can a a high enough density of red-photons be establish for this to happen.Using red light has a second...

  39. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...UV-Violet Dark/Green N/A (high) High 503/ 518 Padron Monomer Blue Dark/ Green N/A (high) Low 503/ 518 rsCherry...specific wavelength of light. PA-FPs can be activated from low fluorescent states to high fluorescence states...fluorescence, to a brighter fluorescent state by exposure to a specific wavelength of light. The switch or...fluorescence when illuminated with 488nm light. However, when UV-violet light is presented first, it causes irreversible...illuminated with violet light, both variants are photoconverted to forms that emit brighter red fluorescence....when exposed to blue light, which is less phototoxic to live tissue than the UV light used by other PA-FPs...State Activating Light Pre/ Post Color Contrast (Fold Change in Fluorescence) Brightness Excitation/ Emission...

  40. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...with exquisite precision using only light. Optogenetics, or light activated tools, can control everything...an experimental optogenetic setup. A light source delivers light pulses to the subject/cells expressing... photoswitches. Light-gated ion channels directly open and close when exposed to light and change the ...changes with light stimulation to either activate or inhibit a specific receptor or pathway.  Light-gated ion...turn on with a light-pulse at one wavelength and stay on until a second pulse of light, with a different...pathways with light. This is achieved by coupling endogenous signaling molecules with light-activated partners... intensities of light sources used to prevent activation of both tools with each light source used.   ...

  41. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...dim" you might get the impression of a candlelight vs. a floodlight. For fluorophore brightness levels,...flow-cytometry-fluochrome-brightness-spillover Excitation Laser Color Very Bright Bright Moderate Dim...Photons of lower wavelengths are higher in energy, while photons of higher wavelengths are lower in energy...important to know the brightness of individual fluorophores. Table 1 describes the brightness of a few commonly...the cytometer. Brighter fluorophores have a stronger signal. Table 1: Degrees of brightness for a few commonly...-Cy7 are "very bright" when they are excited by the yellow/green laser but only "bright" for the blue ...leading to different levels of brightness, depending on the laser light's wavelength. Source: https://www.bdbiosciences.com...

  42. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ... to their brightness and high degree of overlap. Dyes for DNA/RNA staining should have high affinity and...refers to the emission of light by a substance (fluorophore) that has absorbed light or electromagnetic radiation...whereby excitation of blue light will indicate where FITC is bound through green light emission. Key considerations...also vary in their brightness, depending on the respective fluorochrome. Brightness is impacted by both...of findings. If you’re struggling to select the right fluorescent tools, this blog is here to help. Here...ligand to make your selection easier, as well as highlight some recent advancements within this field. Fundamental...wavelength (Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ground state to an...

  43. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...neuroscience. In fact, precise light-based control of biological processes is highly useful in other fields, ...consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength can be ...to low-intensity light, so off-target effects should be limited. Phytochromes are light-responsive systems...harness the power of light, Christopher Voigt’s lab created the first E. coli light-sensitive two-component...construct senses light, and an obligate chromophore allows the system to respond to various light inputs. In...various light inputs, and an obligate chromophore allows the system to respond. In far-red light or dark...exposure to red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system...

  44. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...LACE system for light-activated CRISPR-Cas9 effector, Polstein and Gersbach fused the light-inducible protein...binding affinity with light stimulation2. The Kuhlman lab created two improved Light Inducible Dimers (iLID...of Arch mutants screening for improved brightness. The brightest mutants were then further mutagenized ...enhanced sdChr construct, named CheRiff, exhibits high light sensitivity and fast kinetics. By combining both... weak or high activities (Bert, et al. 2000). To design pXPG, the authors incorporated a high-copy origin...repository, we can't give all of them attention they righlty deserve, but, in this post, we'll provide a small...2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible CRISPR-Cas9 system for endogenous gene...

  45. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...produce the same amount of light from an Fluc or RLuc reaction. In fact, NLuc is bright enough to detect endogenous...require external light sources. Unfortunately, detecting traditional luciferases’ dim light emissions requires...reporters produced bright signals without exogenous excitation and with OgNLuc being the brightest since it had...mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness and improved maturation time compared...Nano-Lanterns. RLuc8 is 5.5-fold brighter than RLuc but NLuc is 30-fold brighter than RLuc8 (6). Susuki, K, ... characteristics: No photoexcitation High Ion specificity High sensitivity Easy expression Gives ratiometric...coelenterzine substrate to the more stable yet brighter furimazine by Promega’s Chemistry group is a good...

  46. Technical Design of a Western Blot

    Type
    Blog Post
    ...have higher specificity, lower cross-reactivity, and are more consistent lot-to-lot. The highest reproducibility... Tris-tricine 150 V, 1–3 hours Good for higher weight proteins Long running time   It’s important...during long-term storage and has both high binding capacity and high protein retention. It comes in a variety...usually preferred because of their higher specific activity, slightly lower cost, and smaller size. However...block with? It can be so overwhelming that you might just stick with the protocol your labmate handed...can help amplify a weak signal. However, they have high cross-reactivity, vary significantly from lot-to-lot... your assay. The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people...

  47. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...ambient light, and versatile as it can be activated by both low and high-intensity blue light. Unlike ...experiments that require high photostability, such as time-lapse imaging or with high light power detection of...Single-cell Phenotypic Observation and Tagging with Light (SPOTlight) as a way to isolate individual cells with...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...here.  Here's what you'll find in this post: Blue-light inducible Cre for use in bacteria CRISPR-Casɸ mGold...New items from the viral service A fast blue-light inducible Cre for precise control of bacterial gene... the use of optogenetics. Optogenetic tools use light to induce conformational changes in a protein.    ...

  48. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ... FPs, which combine high cellular brightness and monomeric state, are highlighted in blue. aExtinction...consider effective brightness of NIR FPs in cells, in addition to their molecular brightness (a product of ...imaging NIR FPs, consider using light sources that produce enough light intensities in NIR. Usually, mercury...with blue-light controlled optogenetics, such as channelrhodopsins and tools based on light-oxygen-voltage...phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than previously reported widely used...Molecular brightness vs. iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa ...Baloban, M. et al. Designing brighter near-infrared fluorescent proteins: insights from structural and biochemical...

  49. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...Signal strength Lower Higher Sensitivity Higher Lower Background Lower Higher Photobleaching/phototoxicity...Luminescence Fluorescence Source of light Chemical reaction High-energy photons Kinetics of photon ...to be extra bright. Other luciferases have been modified to change the wavelength of light they emit or...bioluminescence is the firefly. Along with charming nighttime displays, these insects have contributed an important...reaction in its abdomen to produce yellow-green light (Figure 2). Despite the title of this post, this...includes all bioluminescence, is the production of light by a chemical reaction. In contrast, fluorescence...fluorescence occurs when a molecule is excited by a high-energy photon and re-emits a lower-energy photon; no chemical...

  50. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ... (24-30°C). Brightness: Brightness is a measure of how bright is the emission. Brightness is calculated...is the emission of light by a substance that has absorbed light. The emitted light is at a longer wavelength...proteins are excited by UV light or blue light and their emission is green or red light. For example, T-Sapphire...lose the ability to emit light) after prolonged exposure to excitation light. Photostability can be as... & pHTomato (pH sensors), HyPer (H2O2 sensor), ArcLight (voltage sensor), and iGluSnFr (glutamate sensor...If you do not have a filter that will pass blue light to the detector/camera, then BFPs are of no use ...using more than one FP, make sure their emission light does not overlap in wavelength. In many microscopes...
Showing: 101 - 150 of 986 results