...extensions and, most importantly, any connections the neurons make to other distant brain areas. It is ...axons to their cell bodies. Anterograde and retrograde tracing take advantage of existing transport pathways...J.T., Hantman, A.W., Looger, L.L., Schaffer, D.V., and Karpova, A.Y. (2016). A designer AAV variant permits...implications their signaling is likely to have). Anterograde tracing outlines neurons from their cell bodies...pathways in neurons. Anterograde transport is typically used for the trafficking of organelles, such as ...retrograde transport relies on dynein, while anterograde transport relies on kinesin. Different rates ...of speed for various forms of retrograde and anterograde transport suggest that there are several parallel...

...stimulation guarantees that the majority of the opsins will receive sufficient quantities of photons to...scatter effect is also an important consideration when preparing the implant itself, which brings me to...polished to significant smoothness to prevent drastic loss of efficiency between patch cable and implant. A steady...optogenetic implants have been reported by several groups [4, 5, 6]. These have the obvious advantage of not...opsin through a small piece of fiber optic cable implanted into the animal’s skull (right). The fiber optic...skull using dental cement (a similar procedure as implanting a guide cannula). A fiber optic patch cable is...portion of optogenetics (though it does require a significant amount of equipment, most of which is relatively...

...principal advantage of epitope tags for IHC is the availability of reliable primary antibodies for their...particularly when antibodies to the POI are non-specific, cross-react with other antibodies, or are unavailable...FPs with facile antibody recognition of epitope tags. Thus they have created antigenic protein tags called...their direct fluorescent signals or using anti-FP antibodies. GFP emits light in only a single channel...prone to aggregation and cross-react with anti-GFP antibodies, making them difficult to use in a multicolor...their multiple, highly specific antibody binding sites and primary antibody-based detection, smFPs create...neuronal networks. For almost three decades, recombinant Adeno-associated virus (AAV) has been widely ...

... Pooled library to study antibody escape in SARS-CoV-2 Recombinant antibodies for aminoacyl tRNA synthetases...Validated recombinant antibody toolbox for aminoacyl tRNA synthetases By Kate Harten Recombinant antibodies...Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz As multiple antibody-based...for binding ACE2 is the target of most anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes...binding, but impact recognition by specific anti-SARS-CoV-2 antibodies (Fig 1B). This approach can augment viral...these processes. To create the antibody toolbox, they screened antibody candidates using a synthetic human...whether the generated antibodies could bind these aaRSs. They found the antibodies were able to immunoprecipitate...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...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...

...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...

... 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...

...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...

...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...

...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...

... 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...