We narrowed to 1,082 results for: ANT
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TypeBlog Post...two antibodies against important targets for neurodegeneration research: Anti-MAPT [AT8] and Anti-APP ...doi.org/10.1016/j.cell.2024.01.028. New recombinant antibodies for neurodegeneration research By Ashley...Anti-APP [6E10-A5]. Both are recombinant mouse monoclonal antibodies targeting the human genes and are recommended...the Addgene Antibody Data Hub. Figure 5: Immunohistochemistry with Addgene's Anti-APP [6E10-A5...-A5] recombinant antibody in human postmortem brain samples. Results show positive staining in tissues... Find more neuroscience antibodies here! Wu, Z., Wang, W. (2024). Anti-APP [6E10-A5] in Immunohistochemistry...months, we highlight some of the new plasmids, antibodies, and viral preps in the repository through our...
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Hot Plasmids: Summer 2024
TypeBlog Post...s Neuroscience Antibody Collection to find a recombinant anti-GFAP [N206A/8R] antibody shared by James...well as antibodies for identifying other cell types in the nervous system! Find recombinant anti-GFAP [...editing CHARM: A compact epigenetic silencer Recombinant antibodies for marking astrocytes MagIC-Cryo-EM ...recombinase variants to optimize PASSIGE efficiency (Pandey et al., 2024). The best-performing variant yielded...TALEs), with fifteen total variants deposited with Addgene. These variants, including a “miniCRISPR” version...doi.org/10.1016/j.cell.2021.03.025. Recombinant antibodies for marking cell types in the nervous system...N206A/8R] antibody here! Figure 5: Free-floating rat brain sections stained with Anti-GFAP. The... -
Deep Dive: Fixing and Permeabilizing for Immunofluorescence
TypeBlog Post...Availability Antibodies 101: Introduction to Antibodies Resources on Addgene.org Find recombinant antibodies...recover good antibody binding by tweaking those parameters, but in some cases your antibody may just not... generally referred to as Antigen Retrieval. I chose not to get into antigen retrieval here because it... the Addgene blog: Antibodies 101: Introduction to Immunofluorescence Antibodies 101: Epitope Availability...antibodies at Addgene Find plasmids encoding recombinant antibodies at Addgene ...your samples in a given state or point in time. You want to halt degradative processes while maintaining ...epitope of some targets, preventing your primary antibody from binding (Fig.1). The level of cross-linking... -
27 Hot Plasmids from 2016
TypeBlog Post...expressing plasmid to generate a population of mutant cells. The mutant cells were treated with WNV at a dose ...editing and mutant library generation in several biotechnologically and clinically relevant bacterial species...and CHIP-seq), genome-wide identification and quantification of enhancer activity have remained challenging...sufficient to kill all control cells and surviving “mutant” colonies were deep sequenced to identify the gRNAs...Identifying new ligands for orphan GPCRs is of significant medical interest, however efforts to do so have...gene was chosen to represent the primary splice variant in 38 types of human cancer. Many of the transcripts...vivo expression, and/or produce vectors to make mutant cell lines or transgenic animals. Wall, et al... -
Hot Plasmids Spring 2024
TypeBlog Post...research. Two new antibodies, Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] and Anti-SARS-CoV-2 Nucleocapsid...Addgene. These antibodies are recombinant versions of mouse monoclonal antibodies generated and characterized... for details on the original antibodies. Find Anti-SARS-CoV-2 Antibodies here! Terry, J.S., et al. (2021...38570691. Antibodies to facilitate SARS-CoV-2 research By Ashley Waldron Ready-to-use recombinant antibodies...were immunoblotted against Anti-SARS-CoV-2 Nucleocapsid Protein antibodies #211756 (left panel) or #211757...months we highlight some of the new plasmids, antibodies, and viral preps in our repository through our...pooled CRISPR screens Improved prime editor PE7 Antibodies to facilitate SARS-CoV-2 research New viral vector... -
Healthcare Consulting: A Door to the Business of Life Sciences
TypeBlog Post...context of significant clinical uncertainties and a rapidly evolving healthcare space. The consultants who enjoy...parts of the job. Is consulting meant for PhDs? Healthcare consultants are usually trained in the life...the things that pleasantly surprised me in my first few months as a healthcare consultant was how much my...great fit for former healthcare consultants. I have even seen consultants go back to research and bench ... I’ll share what I’ve learned as a healthcare consultant and give you an idea of what a career in this...skills you’ll need to do well as a healthcare consultant and what you could do to break into the field.... What is healthcare consulting? Healthcare consultants advise pharmaceutical companies, biotech start-ups... -
Hot Plasmids - January 2023
TypeBlog Post...Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder YFP CRISPR-Csm: A...34385696; PMCID: PMC9015817. Meet anti-GFP [N86/8R]! by: Ashley Waldron Anti-GFP [N86/38.1R] was among the...our most commonly requested antibodies. But sometimes, one antibody per target just isn’t enough and we ...second GFP antibody? Similarities: Generated against full length GFP and converted to recombinant form by...reproducibility of a (recombinant) monoclonal. Another reason is that some antibodies may just not perform...section on the Anti-GFP [N86/8R] page for more details. Image credit: Addgene. Find anti-GFP [N86/8R... toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex immunolabeling... -
15 Hot Plasmids from 2017
TypeBlog Post...fluorescent lifetime of 1.3 ns, quantum yield of 0.20). All three mScarlet variants have shown great performance... new RFP variants The toolbox of fluorescent proteins (FP) for cellular imaging is constantly expanding...Campbell’s lab added two new mCherry variants to the repository. These variants can be used as tools to learn... Podcast Segment The impact of KLF4 N-terminal variants on iPSC generation Deriving iPSCs (induced pluripotent...transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered ...Genetically modified organisms like transgenic mice are important tools for understanding the function of cell types...smooth endoplasmic reticulum (OSER) assay. A major advantage of mCyRFP1 is its ability to be coexcited along... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog Post...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 ... -
Hot Plasmids - November 2023
TypeBlog Post...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... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog Post...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... -
To Each HIS Own
TypeBlog Post... 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... -
Selecting Your Plasmid Purification Kit
TypeBlog Post...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... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog Post...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... -
Deep Dive: qPCR
TypeBlog Post...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... -
28 Hot Plasmid Technologies from 2015
TypeBlog Post...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... -
No Llamas Required - Synthetic Nanobodies Against Membrane Proteins
TypeBlog Post... 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... -
A Guide to Starting Your Own Journal Club
TypeBlog Post...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... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog Post... 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... -
Plasmids 101: Shuttle Vectors
TypeBlog Post... 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...