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We narrowed to 884 results for: ANT;

Showing: 121 - 150 of 884 results

  1. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...are a few of the reasons antibodies may result in a lack of response. Antibodies targeting the tumor-augmenting...effectively in mice. Chimeric antigen receptors T cells producing chimeric antigen receptors (CARs) on their... an important element of successful tumor progression. A tumor’s genetic composition is constantly in ...still many important questions in this exciting field of research. One of the most important: How will ...commute to and invade multiple organs, and most importantly escape the immune mechanisms of the host. The...acquire genetic alterations and alter cell-surface antigen production to evade the immune system. 3. Escape...shield themselves from attack. It is a region of incessant chemical activity, and consists of other cell ...

  2. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...versions of Anti-PSD-95 [K28/43R] in rabbit, chicken, and rat. Find Anti-PSD-95 antibodies here! Andrews...reused under CC-BY 4.0 License. Importantly, even commercial antibodies can be tethered on the viral surface...assays like IHC. In fact, Anti-PSD-95 [K28/43R] has been one of the hottest antibodies in Addgene’s collection...Lab’s use of Anti-PSD-95 [K28/43R] under the Application section of the Addgene antibody page, and learn... toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex immunolabeling...months, we highlight a subset of the new plasmids, antibodies, and viral preps in the repository through our...history with protein ticker tape Teaching an old antibody new tricks   New compact, efficient prime editors...

  3. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...2nd mutant strand: The second mutant strand is synthesized in the same manner as the first mutant strand...measure the frequency of each variant during the selection process. Each variant is assigned a functional ...of a plasmid library that expresses all desired variants of a protein. Applying a selective pressure winnows...winnows the pool down to plasmids expressing variants with optimal function. High-throughput DNA sequencing...process is the ability to generate full libraries of mutant proteins. Researchers from the Whitehead lab developed...that can be used to generate complex libraries of mutant plasmids ready for deep mutational scanning.   ...mutational scanning Hundreds of thousands of protein variants can be tested in one experiment. There are many...

  4. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ... 101: Protein Tags Antibodies 101: ChIP Antibodies 101: Introduction to Antibodies References Hochuli ... can also be used as a target for antibody based assays. Antibodies are useful tools for studying proteins... the lab (for a refresher see Antibodies 101: Introduction to Antibodies.) Unfortunately, for some proteins...fused to polyhistidine, then they can use an anti-HIS antibody, like this one developed by Dr. James Trimmer...polyhistidine tags. For more antibody-related information, sign up to receive alerts for antibody posts from our blog...Resources on addgene.org Addgene Antibody Guide Ready-to-use antibodies from Addgene More Resources on ...expressed in frame on the N- or C-terminus of a recombinant protein of interest. The tag was developed as...

  5. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    Published
    May 27, 2025, 1:15 p.m.
    ...and up). But perhaps more importantly, the DNA needs to be free of contaminants, such as bacterial endotoxins...viability. Avoid Low Quality/High Quantity – We recommend avoiding a High Quantity/Low Quality mindset for any...department for these.  While budget is an important consideration, you do want to make sure you’re getting the ...standby to fall back to? We have outlined some important considerations to help you choose the plasmid ...you may need based on their relative quality and quantity.   Figure 1: Plasmid preps fall into different...categories based on their relative qualities and quantities. Image created in BioRender.com. Transformation...Transformation Grade DNA (low quality/low quantity) – Bacterial transformation protocols require such small...

  6. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...four novel SpCas9 variants with altered PAM binding specificity: SpCas9 variant Mutations (relative...SpCas9) PAM sequence D1135E variant D1135E NGG VQR variant D1135V, R1335Q and T1337R NGAN...NGAN or NGNG EQR variant D1135E, R1335Q and T1337R NGAG VRER variant D1135V, G1218R, R1335E...the VQR and VRER variants is similar to wild-type SpCas9, indicating that the variants are likely just ...reported the development of near-PAMless Cas9 variants. These variants, named SpG and SpRY, can target NGN PAMs...the commonly used S. pyogenes Cas9 (3'-NGG) are abundant throughout the human genome, they are not always...limitation: 1) the use of novel S. pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9...

  7. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...execution:  Accuracy is not as important as precision for relative quantification. A consistent 5% positive...up quickly by abundant transcripts, leading to falsely lowered levels for less abundant transcripts. However...qPCR is most commonly used to measure relative quantification of mRNA, which can be used as a proxy measure...between 70-200 bp long With ΔΔCT analysis you’ll want to stay under 150 bp long Annealing temperature...temperature should be 60-65°C For qPCR, the important primer variable is the annealing temperature, not the melting...melting temperature. Though you’ll want to predict this value using BLAST, you’ll need to experimentally...be validated and your reaction optimized. You’ll want to run gradients for optimal annealing temperature...

  8. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...missing codons from the antibiotic resistance gene is inserted. It is important that primers be designed... a library of Arch mutants screening for improved brightness. The brightest mutants were then further ...University have developed two new color variants of their Nano-lantern technology, first developed in 2012...and orange Nano-lantern fusions are designed to complement the original yellow variant, with all 3 having...molecule imaging and inducible gene expression Brilliant technologies adopt basic biological concepts and...principle of protein multimerization is seen in many important biological contexts, such as the enhancement of...Multimerization in the SunTag system occurs via antibody-peptide labeling; specifically, cognate peptide...

  9. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ... targets, similar to how antibodies do. The structure of the common antibody is heftier (~150 kDa) and...fragment derived from a camelid antibody called a heavy chain antibody (HCab). Because of their small ...This step of the protocol starts out with anti-Myc antibodies bound to an ELISA plate to capture Myc-tagged...over several days and isolated special camelid antibodies that bind to the spike proteins in cell culture...especially challenging when trying to develop antibodies against unstable targets like membrane proteins...Walter et al., 2020). No llamas required. The advantages of synthetic nanobodies Nanobodies are small ... highly variable regions that bind to specific antigens. In contrast, nanobodies are a single heavy domain...

  10. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...by both the number of participants and the standard academic calendar. So you want to start a journal club...interested. You don’t want to pick a topic and have no one show up. At Addgene, we wanted to think more closely...away from academic science; there is most likely abundant opportunity to continue such discussions. Beyond...emerging technologies to remain competitive and relevant. Enter: the journal club.  A journal club is ...bioinformatics and HTS, we all had much to learn. And, many wanted to learn more, even those not directly involved... in the project.  Once we gauged interest from relevant parties, and the company at large, by polling ...Preparing for the first meeting There are many important pieces before that first meeting takes place to...

  11. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    Published
    Feb. 8, 2018, 12:17 p.m.
    ... and 70186, respectively). Antares Chu, J., et al. (4), likewise, wanted to produce a BRET probe for ...NLuc (CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into...brighter signal from Antares as compared to Fluc in this model. The construct for Antares is available through... their Nano-Lanterns with NLuc and called their improved creations Enhanced Nano-Lanterns. These great...strategies and tactics for getting the plasmids he wanted. Addgene makes it so much easier to share your ...we share? NanoLuc® Luciferase-based tools are fantastic examples of technologies that can accelerate your...(FLuc) and Renilla (RLuc) luciferase. This is important not so much for how bright you can make a reaction...

  12. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ... tumefaciens, a plant-infectious bacteria that can incorporate foreign DNA into plant genomes. Eukaryotes...contain it. Antibiotic selection Most bacterial plasmids used in the lab contain an antibiotic resistance... you to select for your vector. Some antibiotics block important cellular pathways shared by bacteria ...eukaryotes. But be careful — antibiotic mechanisms vary! Aminoglycoside antibiotics also block protein synthesis...kingdoms of life. That’s convenient for researchers who want to do experiments in complex eukaryotic cells, but...include oris for both target species. Fun fact! Plant vectors are usually designed to shuttle between ... replication site. Regardless of the species you want to be able to shuttle your vector between, you should...

  13. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ...over 13,000 plasmids! Our recombinant anti-c-Myc [9E10] (isotype IgG2a) antibody is currently recommended...the Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody here...light-gated potassium channelrhodopsins Myc-tag antibody anti-c-Myc [9E10] now available! Improved voltage....org/10.1101/2021.09.17.460684.)   Myc-tag antibody anti-c-Myc [9E10] now available! by: Ashley Waldron...months we highlight a subset of the new plasmids, antibodies, and viral preps in the repository through our...something hot? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) here. Here's what...indicators (GEVIs) based on the ASAP sensors. The resultant JEDI-2P sensors overcome many challenges in 2-...

  14. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...expression of recombinant phosphoproteins Protein phosphorylation is one of the most abundant forms of post-translational...while the Patron Lab MoClo Plant Parts kit contains modules specific for plant transformation. Weber et...modules (plant promoter, N-terminal tag, coding sequence of the gene of interest, C-terminal tag, plant terminator...target genome. They did this by selecting the important elements from existing transposon and vector systems...only recently made available through Addgene and we want to highlight it for use in your research. Martínez-García...three dimensional structure of GFP, its brighter mutant S65T-GFP, and the subsequent biochemical characterization...Mitochondrial health, transport and turnover are important features to be monitored to study mitochondrial...

  15. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ... a rigid cell wall makes them resistant to antibiotics and antibacterial drugs like penicillin and streptomycin...Perform culturing without the use of antibiotics since the use of antibiotics can allow mycoplasma to develop...develop resistance to the antibiotics. Antibiotics must only be used to remove mycoplasma and not for preventing... of the quinolone family of antibiotics and are broad-spectrum antibiotic agents. These agents can be ...guest blogger, Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology...largest source among those mentioned above. The contaminants can spread through dirty clothing, lab wears... human scalp, sneezing, coughing, etc. Also, a constant in-flow of individuals wherever the cell cultures...

  16. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...Macaques N/A Stanton et al., 2023 AAV-MDV1A CNS Mice No (LY6C1) Stanton et al., 2023 ... way that goes beyond such restrictions, you may want to consider a systemic capsid for your AAV.   Systemic...unique tropisms are now available. These include variants with broad CNS or PNS transduction, cell type ...concurrently to minimize the impact of neutralizing antibodies developed in response to the first exposure to...determine the best titer for your application.   Pro tip!Want to know if a capsid is a good option for your experiment...effective in both mouse strains. It is therefore important to review the literature for use of a capsid in... brain endothelial cells in mice but neurons in infant macaques (Chen et al., 2023). And these differences...

  17. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...reverse transcriptase mutant in a separate control reaction. This will allow you to quantify the amount of PCR...viral prep, treated with the antibiotic, and colony-forming units quantified. While these methods tend to...of a gene carried on the transfer plasmid or quantifying the number of viral copies that have integrated...commercially available p24 ELISA kits that can quantify p24 levels in a matter of hours. A major drawback...particle. Consequently, titers based off of p24 quantification tend to be overestimates since they can include...viral RNA is first converted to cDNA and then quantified using qPCR primers targeting specific viral components... components such as LTRs, gag, WPRE, antibiotic resistance-genes, or the transgene itself. Many researchers...

  18. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    Published
    March 7, 2023, 2:15 p.m.
    ...synergistically, antagonistically, were additive, etc.). ANCOVA and MANCOVA All variants of ANOVAs discussed...there is a statistically significant difference in that variance. If you want to know type of distribution...normality (or lack thereof). Alternatively, a Quantile vs. Quantile plot can determine if your data deviates...sets. Significance is a numerical value which quantifies the likelihood that a result is due to chance.... Below are several core factors of statistics relevant for many downstream statistical tests.   Independent... Variance – Approximately equal Data type - Quantitative If your data doesn’t fit the requirements above...assess if the mean of an experimental group is significantly greater or lesser than the mean of a control...

  19. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...multicopy variant of a gene exists, there must be assays that researchers can use to quantify said copies...ddPCR provides absolute quantitative data, instead of relative quantification.  Even though no one option...: 15833050. Ma L, Chung WK. Quantitative analysis of copy number variants based on real-time LightCycler...PCR-based applications (qPCR and ddPCR.) qPCR Quantitative/real-time PCR (qPCR) has been used for copy ...expressed, one-copy ‘housekeeping’ gene), and quantified relative to a 'normal' sample with known copy...about qPCR is that since it measures relative quantification - that is, since you’re looking at the amount... dye, with a fluorescent probe and quencher to quantify specific amplicons.  Sybr has high background ...

  20. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...activity (Ni, Titov, & Zhang, 2006), quantifying antibody/antigen interactions (Saraheimo et al., 2013...contributed by guest blogger James D. Fessenden, an Assistant Professor at Brigham and Women’s Hospital. Biochemists... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...nonspecifically soak up your fluorophore package. One important concept in FRET experimental design is that the...donor first where labeling specificity is most important. Donor labeling and the need for specificity Donor...protein molecules contain the donor and, more importantly there is no fluorescence background resulting...genetically-encoded FRET donors (Fig. 1). Scores of FP variants have been created, thereby enabling a broad spectrum...

  21. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...while the anti-HA frankenbodies include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody...determining region of an anti-FLAG scFv and a stable scFv scaffold. (b) Schematic of anti-FLAG-FB-GFP binding...localization. Adapted from Liu et al. 2021. Anti-FLAG and anti-HA frankenbodies have a variety of applications...up here.  Here's what you'll find in this post: Anti-FLAG Frankenbodies for modular imaging of cellular...system for mammalian genome regulation and editing Anti-FLAG Frankenbodies for modular imaging of cellular...cellular targets by: Mike Lacy Adding to their set of anti-HA frankenbodies (Zhao et al. 2019), Tim Stasevich...Hiroshi Kimura’s labs have recently published a set of anti-FLAG frankenbodies, a flexible, modular tool for...

  22. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...asked questions. We also want to tell you about things you didn’t know you wanted to know about, like research... the example of our Antibody Guide, which was published alongside our new antibody services in the spring...While Rachel had antibody experience, we also had the deeper expertise of our antibody team to rely on....In the case of the antibody guide, primary research and previously written Antibody 101 blog posts were... statement extends beyond plasmids, AAVs, and antibodies to our content, website, Data Hub, and everything...that process. Our repository of plasmids, AAVs, antibodies, and data allows scientists to easily and quickly... series of conversations on resources with our antibody and content team, we identified what applications...

  23. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...major advantage of using this method for lambda red expression is that it doesn’t require antibiotic selection...  References 1. Costantino N, Court DL. Enhanced levels of λ Red-mediated recombinants in mismatch repair...outgrowth of bacteria. Selection and confirmation of recombinant clones.    Figure 2: Overview of using... system to replace a gene of interest with an antibiotic resistance cassette.   Substrate DNA Design..., including selectable DNA fragments, such as antibiotic resistance genes, as well as non-selectable DNA...replacements and tags.  The typical frequency of recombinants is 1 positive clone out of 104 to 105 colonies...

  24. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    Published
    June 8, 2021, 1:15 p.m.
    ...can vary "Lab assistants" usually wash glassware, prep media, etc. "Research assistants" are more involved...daunting. During my first year of college, I knew I wanted to try research, but had no idea what to expect...investigator (PI) Runs the lab Writes a lot of grants Varying degrees of engagement with what happens...the lab Official title is typically Professor, Assistant Professor, Associate Professor Sometimes teaches...independent project. Lab technician / Research assistant Typically has a Master's or Bachelor's degree...order materials, lab maintenance Administrative assistant: assists PI with admin duties Staff scientist:...worthwhile and can help us make decisions about what we want to do in the future. How do I get started with research...

  25. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ...utilization, Arabidopsis has made significant contributions to understanding plant and crop science alike.  Arabidopsis...for use in Arabidopsis. There are also important tips for plant transformations and the use of Arabidopsis... The development of Arabidopsis as a model plant. The Plant Journal, 61(6), 909–921. https://doi.org/10.1111...page Find more plasmids and resources for plant resources on Plant Plasmids Collection Page Read our molecular...Justin Ng and Alyssa Cecchetelli Science is a constantly evolving and demanding field, requiring a variety...vastly expanded axolotl research. One of the most significant advances is the sequencing of the entire axolotl...research providing insight into some of the most significant and fundamental biological processes. Many of...

  26. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...site-directed mutagenesis.  Quantitative polymerase chain reaction, qPCR Quantitative polymerase chain reaction... to quantify the concentrations of multiple starting DNA templates in one sample. It is important to note...heating and cooling steps. PCR relies on heat resistant DNA polymerase from the thermophilic bacterium...aquaticus (Taq). Taq polymerase is thus a heat resistant enzyme that can withstand changes in temperature...to confirm product presence, size, and relative quantity. End point PCR is most commonly used in molecular...genotyping. It’s extremely useful but is not as quantitative as other methods of PCR. Theoretically scientists...scientists should be able to determine the quantity of DNA after a PCR reaction as the amplicon doubles every...

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

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ...substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby excitation...agonists and antagonists, impacting the interpretation of experimental results. It's important to consider...required. These dyes can also be attached to an antibody to detect various targets, such as proteins, nucleic... These consist of a genetically encoded enzyme variant tag that covalently reacts with a small substrate...through fluorescent labeling. There are several advantages to using these self-labeling proteins over traditional...near-infrared fluorophores. Fluorescence microscopy takes advantage of the characteristics of these fluorophores to...protein function and cellular components: It's important to assess how dye-conjugation affects protein ...

  28. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ... you want to use optogenetics The most important question to consider is what exactly do you want to use...and components necessary to run it? And, most importantly, what are the methodological and experimental...considerations? I recently spent two months in the lab of Antonello Bonci, Director of the National Institutes of ...optogenetics for? The technique is non-trivial; you don’t want to spend months doing surgeries and making fiber...region you wish to stimulate 2) Fabrication and implantation of the optical fiber into the region of interest...any optogenetic experiment. There are numerous variants for the different types of opsins available and...of North Carolina) that offer his constructs. Importantly, control viruses (not expressing the opsin) are...

  29. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...plasmids, multiple Cas9 variants are available: wt humanized Cas9, D10A nickase mutant (Cas9n), dCas9 (transcriptional...pCAMBIA. Cas9 is plant-optimized, and plasmids are available for both monocot and dicot plants, with a choice...Cas9 toolkit for multiplex genome editing in plants.” BMC Plant Biology 14 (2014): 327. PubMed PMID: 25432517...multiplex genome editing in monocot and dicot plants.” Molecular Plant 8(8) (2015): 1274-1284. PubMed PMID: 25917172...suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!   Why use multiplexed...This increases the chance that all the edits you want to make will happen.  There are many reasons to ...with Golden Gate and Gibson assembly methods If you want to scale up the number of gRNAs in your plasmid,...

  30. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...expressing the recombinant plasmid to propagate. Any cells that contained non-recombinant vectors (re-ligated...DB3.1, which contains a mutant version of DNA gyrase (gyrA462) that is resistant to the toxic effects of...factor plasmid of E. coli, is part of a toxin-antitoxin system encoded by the ccd operon, which is responsible...another gene found in the ccd operon, codes for the antitoxin protein (CcdA) that protects the cell against ...the number of clones that do not contain the recombinant plasmid and therefore makes the cloning process...modern version of this older system, with added advantages that will be discussed in detail in a separate...post. Focusing on the ccdB aspect, Gateway takes advantage of the same principle that cells will not propagate...
Showing: 121 - 150 of 884 results