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We narrowed to 987 results for: IGH@;

Showing: 401 - 440 of 987 results

  1. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...Immunofluorescence (IF), is an immunoassay that brings to light the cellular world. The technique allows you to ...indicates your target’s location. Some other terms you might see associated with IF are immunocytochemistry (... of other antibodies. But, once you do find the right protocol, multiplex IF can give you impactful and...sample on slides, and confocal   Figure 2: High-level overview of an indirect immunofluorescence...ones? Choosing a secondary is often much more straightforward than choosing a primary, so we’ll start there...the future.  We hope that this post has shed some light on immunofluorescence. The technique can provide...

  2. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...Every few months we highlight some of the new plasmids, antibodies, and viral preps in our repository...microscopy (cryo-EM) protocols typically require high concentration and purity of the target molecule,...or hundreds of particles surrounding each of the highly-visible beads on the cryo-EM grid.     ...denaturated (left) or complexes may dissociate  (right). B) Protein samples protected from AWI by helical...authors to determine structures at comparable or higher resolution than using more complex protective methods...

  3. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    ...heterodimers composed of one of 18 alpha and one of eight beta subunits. Both subunits have short intracellular...ectodomains can extend and open, binding ligands with high affinity. This extended-open state is key to strengthening...flexibility is crucial for their function. With the right research reagents, researchers can measure this ...developed and defined. According to the Springer lab, a highly specific function-blocking antibody, αVβ3.13, may...as new reproducible rat or mouse rAbs. The other eight are chimeric antibodies composed of rat or mouse... recently showed RGD-binding integrins demanded higher tensions before they would enable cells to spread...

  4. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...kDa single polypeptides that contain the variable light chain (VL) and variable heavy chain (VH) of an antibody...minimizing oligomerization of the scFv. While you might assume that the variable domains of an scFv would...lower affinities, lower long-term stability, and a higher likelihood to aggregate due to their small size...library with plate-bound antigen helps select for high affinity scFvs. Bound phage are then proliferated...with plate-bound antigen to select for scFvs with high affinity binding. Bound phage are then proliferated... since direction injection to the eye results in high drug concentrations in the eye with minimal systemic...

  5. Editor's Choice, July 2016

    Type
    Blog Post
    ... To better highlight the great content contributed by our bloggers each and every month, we've decided...Cpf1 Update: Comparison to Cas9 and NgAgo As you might expect given the excitement surrounding anything...month is from guest blogger James D. Fessenden at Brigham and Women's Hospital. This post describes how to... our posts if you have any questions or useful insights. If you're interested in blogging for us, fill...

  6. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...fortunate that there’s a guide that directs cleavage right before the stop codon!  Figure 3: How to ...guide sequences (arrows) and their PAM (bold highlighted text) and cleavage sites (slashes) are shown....plasmid options for templates - see the example at right using blue flame plasmid pmScarlet_C1! After creating...primers to add T7 promoter-left homology arm and right homology arm to the desired insert (e.g., mScarlet...the main drawback of i-GONAD is that it requires a high-concentration, column-purified DNA template.   ...

  7. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...not having any confounding mutations, the risk is high. You are generating a gene therapy product with ...clinical trials. An off-target rate of 5% is far too high when considering human patients and the diseases...measurements at those sites to get an idea of how high the off-target frequency is (more on this below)...sequencing methods. Candidate site sequencing Sites with high sequence similarity to the gRNA can be sequenced...a DNA mutation, off-targets must be quantified slightly differently. For a comprehensive approach, RNA-seq...library screens, the same principle holds true - the higher the number of distinct editing events, the lower...

  8. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...Every few months we highlight some of the new plasmids, antibodies, and viral preps in the repository...deactivation). Both have significantly improved brightness, kinetics, and sensitivity. iGluSnFR4f is well-suited...and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's what makes these new backbones...Protein Innovation (IPI) recently released a set of highly-specific recombinant antibodies against three of... — with efficiencies between 5% and 11%. It has higher efficiency and lower indel formation than methods...

  9. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...fixed sample. Depending on the sample type, this might be frozen or formalin-fixed, paraffin-embedded tissue...Depending on your needs, your sample, and targets, you might decide to do simultaneous IF instead (Figure 1B).... Depending on the goal of your experiment, you might need to quantify the intensity of the different ...spatial relationship between the targets. Or, it might be sufficient to describe or classify the labeled...label a sample that expresses your target(s) at a high level, either naturally or by overexpression, for...similar between the two experiments. If not, it might be that the other antibodies are disrupting the ...

  10. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...vectors containing high numbers of gRNAs in one overnight reaction. STAgR is highly customizable as vector...In this quarterly blog series, we’ll highlight a few of the new CRISPR plasmids available at Addgene....

  11. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    ...RNA is highly desirable for extending the complexity of genetic circuits, allowing for tighter spatio-...Illinois at Urbana-Champaign. The PUF proteins include eight modular repeats, each of which binds a consecutive..., I think, are endless.” Abil says the new tool might even find therapeutic application in addition to...she recommends using standard BsaI in place of the high fidelity version (despite what the original paper...

  12. Scientist Networking: What is an Informational Interview?

    Type
    Blog Post
    ...connection to the hiring manager. They might say yes and they might say no but do not confuse networking...scientist in the academic system has many pluses. I delighted in my graduate school years for allowing me to...personal questions can elicit the most valuable insights you will get from the interaction. Most people...great way to practice for real interviews where you might spend a whole day having to talk with strangers....

  13. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ... all night was hand-picked (like our plasmid samples!) by Addgenies. If you were there, you might have...plenty of delicious desserts. We enjoyed a video highlighting the changes at Addgene over the last 10 years...Perhaps the most heartfelt portion of the entire night was when Melina and Ken called their parents up ...

  14. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
    ...customized to highlight different skills and to get the important information in the right place on the...organization is small, any person at the company might be able to help you. If the organization is large...additioanal advice for the science career search.  You might also enjoy the other posts in the Addgene Blog Scientist...

  15. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ... homozygous knock-in animals can be obtained in eight days. The only animals on a plate, other than the... delivery, screening, and more. Here are some highlights from the past few years:  Repair template design...improvements from the Mello lab resulted in strikingly high editing efficiencies, obtaining as many as 100 independent...corresponding to the same strand as the PAM produced a higher editing efficiency, while for edits 3’ to the PAM...templates. They found that Cas9 RNPs could be toxic at high concentrations, and editing efficiency improved ...

  16. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...and can exhibit recombination levels as high as 1-3%. In light of this finding, we now include guidance...leaky expression. On the right non-homologous scrambled recombination sights lead to reduced reversion...strategy should work, these vectors suffered from high levels of spurious expression.  As such, researchers...weak and may require longer waiting periods and/or higher injection titers to achieve adequate expression...

  17. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...transposons evolve under contrary selection pressures: high efficiency helps them spread, but also stresses ... evoCAST In the emerging world of gene therapy, highly tailored treatments to specific mutations require...long sequences. In their experiments, PASSIGE had higher efficiency, but evoCAST required less optimization...optimization for new targets and produced a higher-purity product, likely because the integration occurs in a... can’t wait to see where evoCAST takes us on the high seas of gene editing. Bon voyage!   References...

  18. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...proximity to an endogenous gene. Each trap uses a slightly different approach (described in detail below)...for the endogenous gene – its expression levels (high/low) and patterns (tissue/timing) can be monitored...screened. Depending on the marker, traps can be used as high-throughput screens to introduce reporters throughout...the top trap – they don’t require insertion in a highly specific gene region and they don’t disrupt the...Resources References Wood, V., et al., Hidden in plain sight: what remains to be discovered in the eukaryotic...

  19. Summer SciComm Series: Master of SciComm

    Type
    Blog Post
    ...yet familiar. The idea that my piecemeal degree might one day foster a meaningful day job only came into...enjoyed a taste of what was to offer, continued straight on to a master's degree in science communication... one day required of me for work, I would have a high level of career satisfaction indeed.   For many ...science. Understanding just how the public utilizes insights gleaned from scientific research is, after all...law to poetry. A research discipline in its own right, science communication reflects a necessary confluence...

  20. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ... TX-TL system is capable of robust expression of high G+C content genes, as validated through monitoring...CRISPR Plasmids and Resources page. Here are some highlights from the past couple months including several...

  21. Viral Vectors 101: Transductions

    Type
    Blog Post
    ...are very valuable biological tools, and with the right safety considerations, they aren't dangerous for...transductions can offer benefits such as tissue-specificity, high efficiency, and can be selected to be either integrating...our viral selections to see if we have one that might work for your next experiment. Using transduction...Answering these questions will help guide you to the right method for your experiment.  A cool feature of viral...virus can sound daunting, but it is easier than you might think. We have protocols available to get you started...

  22. What the HEK?

    Type
    Blog Post
    ...an advantage to use the cell line and we will highlight some of those applications here.   Virus production...transfect in bulk and produce recombinant proteins at high levels. In a case where clinical applications of...and if your research doesn’t allow for that, HEK might be the cell line for you. Are HEKs for you?   If...you are looking for a human cell line model, HEKs might be a great option! Especially if your research interests...

  23. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...these visualization tools is CRISPR-Sirius, the brightest CRISPR-based tool to date for visualizing genomic...chromosomes in living cells, the system requires high-copy chromosome-specific loci in the genome and ...the gRNA target site. In addition to increased brightness, they also found that the signal to noise ratio...its predecessor CRISPRainbow, CRISPR-Sirius is brighter making it suitable for observing low-copy genomic...copy genes. If you want to visualize genomic loci brightly, give CRISPR-Sirius a try. Find CRISPR-Sirius ...

  24. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    ...with a large range of characteristics (color, brightness, photostability, maturation, oligomerization)...choosing a FP, but much of the primary data one might require when making such a decision remains spread...there is a great deal more information that one might like to store about FPs than is currently represented...multi-parametric search queries.  Looking for a bright, monomeric FP published in the last few years with... for me? A Practical Approach to Choosing the B(right)est Fluorescent Protein Choosing Your Fluorescent...

  25. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    ...grandparents’ medical textbooks by torchlight under the covers. In high school, I went to all the optional...technical jargon that even scientists in neighbouring disciplines might struggle to understand. Why should ...whispered questions from friends and classmates back in high school, and it’s oddly fitting that I’ve ended up...

  26. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...optimal for experiments requiring high signal to noise ratios, imaging of high frequency glutamate signaling...to create SF-iGluSnFR. This improved variant has higher expression levels in bacteria and produces stronger...properties The improved variants of iGluSnFR3 have higher expression levels and produce stronger fluorescent...Biosensors A Practical Approach to Choosing the B(right)est Fluorescent Protein Visit the Fluorescent Protein...

  27. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    ... be ideal for you. Here, we will chat about who might want to explore this role, how to seek out such ...commitment, planning, and funding – they may not be right or necessary for many scientists! Unless some of...from third parties or your home institute. Discuss rights Do you expect to generate data that will contribute...It is never too early to discuss authorship and rights for anything that could be generated during the...opportunity, but also a really good time! If it sounds right for you, start thinking outside of your current ...

  28. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...buck - it grows at higher cell densities than most of its counterparts, yielding higher production power... phaffii in the 1970s with the goal of producing high protein animal feed. Due to increasing prices of...flask volume. What’s even better? K. phaffii is highly efficient at protein secretion, making harvesting...knock-in or knock-out) is essential to your work, you might want to pick a yeast strain best suited to it. Putting...

  29. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ...figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...putting the operon under a strong promoter results in high R-body expression  (Schrallhammer et al 2012). This...environmental conditions. More speculatively, R bodies might endow divergent organisms with new functions. For...understanding of how R bodies assemble and function, we might be able to endow cells with new ways to interact...

  30. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ...sides of the break, we determined that Cas9 held tightly to three of the four strands of the cleaved duplex... the PAM-proximal side, consistently supported higher frequencies (up to 60%!) of gene editing than single...have shown that this technique supports extremely high frequencies of genome cutting and, when donor DNA...included in the nucleofection reaction, extremely high frequencies of HDR. The detailed protocol for nucleofection...Central PMCID: PMC4383097. 3. Kim, Sojung, et al. "Highly efficient RNA-guided genome editing in human cells...

  31. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...In a big lab, the advisor might not be around much and your real boss might be a postdoc – this isn't ...competitive style to spur you to succeed or you might enjoy working closely with others to accomplish ...before you take the position. Am I choosing the right mentor? Make sure your advisors will be good role...candidate to ask lab heads, lab members and lab neighbors during the interview process. What if I want...

  32. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... you to select cells with high GFP expression. These cells have the highest levels of Cas9 and gRNA expression... which method you use, the procedure will vary slightly.   Two Golden Gate options that are available...expression, and thus the highest frequency of genome editing events. Yamamoto Lab Multiplex CRISPR/Cas9.... “Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing.”  Nature Biotechnology...al. “A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and...

  33. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    .... If submitting plasmid(s) using a spreadsheet (right), click the “Download Deposit Spreadsheet” button..., and plasmid description/experimental purpose (right). On the article page, enter the name, type, and...The more sequence data available, the better. We highly encourage you to upload full sequence data whenever...cannot be propagated in DH5α. For plasmids with highly repetitive sequences (which can be prone to recombination...Figure 12: The "Copy" button is located near the top right of each data entry page. Once you have completed...necessary by my institution or other third party rights holders. Once this process is complete, you will...

  34. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...sensitivity. Wild-type Cas9 exhibits high on-target activity, but also high off-target activity, which may ...sensitivity is the measure of on-target activity (right). Cas9 nucleases that exhibit enhanced site ...et al., 2018) and found that Sniper-Cas9 showed a high degree of both specificity and sensitivity across...Evolved Cas9 variants with broad PAM compatibility and high DNA specificity." Nature 556.7699 (2018): 57. PubMed...

  35. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...RNA without altering the genome, but Cas13s have higher on-target specificity. New work from Konermann ...package in low-capacity vectors! These small, but mighty type VI-D enzymes are the latest tools in the transcriptome...including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are described...Cas13d deposit here). Konermann et al.: CasRx is a highly potent, specific editor in mammalian cells Not ...characterized Cas13 enzymes, CasRx also displays very high on-target efficiency; where shRNA treatment produced...

  36. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...is building the software tools for the right people at the right time. To explain a bit further, a software... a strong background of undergraduate research straight to a lab career at a biotech company, I always...operations software for next-gen sequencing (NGS), and high-throughput screening. While some developers had ...about what a software product manager does, you might be wondering what the day to day looks like. Check...

  37. Hot Plasmids - May 2022

    Type
    Blog Post
    ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...close proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids are...mammalian cells. Long sequences traditionally have higher risks of off-target mutations. These plasmids contain...

  38. Hot Plasmids - February 2022

    Type
    Blog Post
    ...to induce high levels of gene activation for both reporter and endogenous genes. It is highly specific ...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...target only requires a small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the...

  39. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    ...hybridoma, you can choose one that produces high-affinity/high-avidity antibodies. The same hybridoma is...drive can be identified by sequencing the heavy and light chains. To circumvent any possible genetic drift...  Figure 2: Changes to the heavy chain or light chain genes within a hybridoma impacts the specificity...validating monoclonal antibodies, their cost is higher than that of polyclonal antibodies. For example...

  40. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...mixture of products or from incomplete PCR — although higher DNA concentrations are needed for this method to...retain products with single-stranded overhangs (right branch). The mixed PCR and incomplete PCR methods...with two 5′ overhangs, so these approaches require higher concentrations of insert to succeed. 5′ overhangs... showed that a five-piece assembly reaction was highly efficient (~80%) with 40-bp homology regions. Ten-fragment...method also based on homologous recombination. The higher temperature at which Gibson assembly takes place...
Showing: 401 - 440 of 987 results