...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...

...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...

... 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...

...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-...

...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...

... 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...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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...

...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...