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We narrowed to 920 results for: Gene

Showing: 521 - 560 of 920 results

  1. A Deep Dive into BioBuilder

    Type
    Blog Post
    Published
    Oct. 11, 2017, 1:44 p.m.
    ...Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ... education policy. Additional Resources on the Addgene Blog Synthetic Biology and the Importance of Assembly...All Synthetic Biology Blog Posts Resources on Addgene.org Guide to SynBio Assembly Standards Find Plasmids...

  2. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...at East Tennessee State University. He works with gene expression in Pseudomonas aeruginosa. Follow him...Protocols Check Out How Addgene Develops Lab Management Software Resources on Addgene.org Find Protoctols ... and do not necessarily represent the views of Addgene. I think it’s safe to assume that anyone reading...by New England BioLabs In my lab we do a lot of genetic construction and cloning experiments. Not all of...100percentSDS.     Additional Resources on the Addgene Blog Learn How to Keep a Lab Notebook for Bioinformatic...

  3. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...endonucleases. A given prokaryote typically has genes encoding one or a few restriction modification systems...Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Check Out Our Plasmid...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation (there are...illustrates how sequence visualization software (we used Snapgene here) can help you assess whether your restriction...describing dam-/dcm- E. coli Resources on the Addgene Blog Keep up with the Latest Cloning Techniques...

  4. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...conducted a CRISPRi screen to identify endogenous genes that impact CGBE efficiency. In addition to using...review how CRISPR-derived base editing works in general. Base editors use either catalytically dead Cas9...efficiency and produced fewer indels than first-generation editors. This team also demonstrated that different...sequences) (Huang et al., 2024). BE4max, a CBE, generated similar deletions at a lower frequency. Their ...and its inclusion significantly reduced indels generated by CGBEs. What's next? Base transversion editing...base transition editing, we can expect several generations of optimization before the field begins to converge...

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

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...In this case, Konermann et al. targeted MAPT, the gene encoding dementia-associated tau, delivering dCasRx...possibilities. To identify Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR ...scientific sharing will continue to grow, and we at Addgene appreciate our depositors for making their tools...29551272 Plasmids from this paper are available at Addgene. Yan, Winston X., et al. “Cas13d Is a Compact ...29551514 Plasmids from this paper are available at Addgene. Learn more about research at Arbor Biotechnologies...Biotechnologies. Additional Resources on the Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips...Cas13a (C2c2) Characterization Resources on Addgene.org: Find RNA Knockdown Plasmids Check out our CRISPR...

  6. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...PMCID: PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which... tagging strategies Additional Resources on Addgene.org Fluorescent protein plasmids Use FRET to study...protein-protein interactions Deposit plasmids with Addgene! ...

  7. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ... widely available and applied by researchers for gene targeting. Fast forward a decade, and the CRISPR...Resources on Addgene.org CRISPR software and resources Addgene CRISPR Guide Addgene’s CRISPR 101 eBook... trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its target in cis and cleaves...end goal. CasPEDIA also features useful links to Addgene resources throughout the site. In the Experimental...each enzyme page, you'll find a link to search Addgene's collection for plasmids containing that enzyme... from the article if they're available through Addgene. So whether you're looking for the exact Cas9 plasmid... or browsing for the latest AsCas12a plasmids, Addgene and CasPEDIA have got you covered!     ...

  8. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ...where we could incorporate cold-tolerance inducing genes within a regular mesophilic host system. This would...resources on the Addgene blog Find our synthetic biology blog posts Resources on Addgene.org Find plasmids... pC013-Twinstrep-SUMO-huLwCas13a plasmids from Addgene, which would potentially allow a stable Cas expression...” by Benoit Giquel, an outreach scientist from Addgene. The talk highlighted the importance of reagent...

  9. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...repair of Cas9-generated breaks is useful if the intent is to make a knockout in your gene of interest, ..., as it is prone to generating indel errors. Indel errors generated in the course of repair by NHEJ are...are typically small (1-10 bp) but extremely heterogeneous. There is consequently about a two-thirds chance...Of some importance, the deletion can be less heterogeneous when constrained by sequence identities in flanking...overhangs is available. Cas9 can also be altered to generate a targeted single strand break; when two such ...  Dale Ramsden is a member of the curriculum in Genetics and Molecular Biology, the Dept. of Biochemistry... Break Diminishes Its Mutagenic Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen...

  10. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...downstream application such as quantitative PCR to assess gene expression. There are several things that researchers...everything, including your skin. Here are some general tips for working with RNA, even if you are using...10.1038/nprot.2006.83 Additional resources on the Addgene blog Read blog posts related to plasmid protocols...for purifying DNA without a kit Resources on Addgene.org Check out our molecular biology protocols Find...

  11. Transferable Skills Guide: Public Speaking

    Type
    Blog Post
    Published
    April 20, 2023, 4 p.m.
    ...You realize, much too late, that you labeled your gene of interest incorrectly on slides 8-16. Your notes...remember related to this paper and/or biology in general until I reached Fig. 6, Panel L, and hope nobody...breath–you got this!  Additional resources on the Addgene blog Transferable Skills Guide: Time Management...

  12. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ... useful as a mechanism of transient knockdown of gene expression. This system could be used in lieu of...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9 has been...Cas12’s primary difference from Cas9 is that it generates staggered breaks, also known as sticky ends. Cas12a...targeting experiments requiring single strand DNA generation, such as homology directed-repair. Cas12a is ... of DNA. This makes it particularly useful in generating transient edits of signaling molecules (RNA) ...If your genome engineering goals expand beyond generating frameshift mutations or single base pair edits...s41586-021-03886-5 Additional resources on the Addgene Blog CRISPR 101: Which Cas9 do I choose for ...

  13. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...Chemical Modifications of CRISPR RNAs to improve Gene-Editing Activity and Specificity. JACS 144, 12584...resources on addgene.org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources...effectively serving their function of guiding Cas9 to generate a targeted break. These advances in gRNA technology...or immunity itself is being studied. Check out Addgene’s validated gRNA repository to find pre-tested gRNA...photocleavable gRNAs. Photocleavable guides are generated by introduction of a single photocleavable 2-nitrobenzyl...lentiviral plasmids Additional resources on the Addgene Blog  How to design your CRISPR gRNA CRISPR 101...

  14. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...from Jiazhang Lian’s lab that enables simultaneous gene activation, repression, and editing with a single...studies.    Duong et al., Genetics 2021. https://doi.org/10.1093/genetics/iyab006       The CRISPR ... need. Here's what you'll find in this post: Epigenetic engineering with single-pot assembly Tracking...CRISPR plasmids New items from the viral service Epigenetic engineering with single-pot assembly By Eric ... of plasmids to the repository that will help epigenetic researchers custom design their own engineering...) Golden Gate assembly (backbones available at Addgene) with examples of potential synthetic chromatin...with BFP, sender cells expressing surface BFP generated BFP-labeled receiver cells. What’s more, G-baToN...

  15. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...have been modified for genome editing, activating gene expression, visualizing genomic loci, and much more...plasmids from this publication at Addgene. Resources on the Addgene Blog Read about Cas9 Homolog Cpf1...effect on specificity was similar; SpCas9-HF1 generated fewer off-target cuts when compared to WT SpCas9...collaboration with the Joung Lab, they used targeted mutagenesis to create HypaCas9 (SpCas9 N692A/M694A/Q695A/...-HF1. The HypaCas9 plasmids are available from Addgene here! Future possibilities for Cas proteins With...26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal structure...26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading governs...

  16. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...cells)  that allow for continuous expression of genes integrated into the insect cell genome. Both of ...and cell free expression systems. Overall the general strategy for protein expression consists of transfecting...systems Yeast are a great expression system to generate large quantities of recombinant eukaryotic proteins... species due to its use as a model organism in genetics and biochemistry. When using S. cerevisiae, researchers...large-scale, stable production of proteins. In general yeast expression systems are easier and cheaper... use. Alyssa D. Cecchetelli is a Scientist at Addgene. She received her PhD from Northeastern University...proteins in plant root exudates Resources on the Addgene Blog Learn about inducible promoters Learn about...

  17. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...fewer transcripts of their antibiotic resistance genes and can therefore be cultured in media containing...plasmid prepping is a breeze. You get your stab from Addgene, streak for single colonies, sub-culture, and prep...use a different strain. For most applications, Addgene uses and recommends DH5𝝰! This strain consistently...density and plasmid yield as these factors are generally, but not perfectly, correlated. Typical growth...sufficient gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around... strain instructions from the manufacturer. On Addgene plasmid pages, you may see depositor recommendations...writing of this post. Additional Resources on the Addgene Blog Visit our Plasmids 101 Topic Page Learn How...

  18. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...error than normalizing to so-called housekeeping genes (Aldridge et al., 2008).  Woah! That was a lot of...quantification Normalizing to ubiquitously expressed genes Not as accurate; increased antibody costs; increased...Antibodies Resources on Addgene.org Addgene's Western Blot Protocol Addgene's Western Blot Protocol Video...Looking for a western blot protocol? Check out Addgene's western blot protocol and protocol video!  Antibodies...gel. Handcast gels are significantly cheaper and generate less waste. However, they also add time to an ...in size. However, these systems are expensive, generate plastic waste, and limit the amount of optimization... can only compare samples to a standard curve generated on the same blot. If your protein of interest ...

  19. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    Published
    July 9, 2019, 12:52 p.m.
    ... terms of the vocabulary--DNA, protein, plasmid, gene, etc. Though I talked about CRISPR’s molecular mechanism...quick tips to get things going: If your PTA has a general meeting and a means to request funding for events...7th grade biology teacher. I led up to my job at Addgene, which is likely a career path none of them had...offered engaging examples of how understanding genetics is so timely.” Genome engineering, and many other...guidance and feedback. Additional resources on the Addgene blog Browse our CRISPR blog posts Download our...science communication blog posts Resources on Addgene.org Check out our comprehensive CRISPR guide Find...

  20. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...Calabrese (activation), from Addgene to better understand which human genes are important for SARS-CoV-...deposited at Addgene. Plasmids deposited from Alejandro Balazs’s lab includes those for generating pseudovirus... deposited the plasmids used for this study at Addgene which include a C-terminal deletion in the spike... Bst polymerase constructs from their assay to Addgene.  Drew Endy’s and Philippa Marrack’s labs also ...investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids express these and other...the COVID-19 pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open ...10.1089/crispr.2020.0030 Additional resources on the Addgene blog Learn more about mapping the SARS-CoV-2 -...

  21. How to Write a Scientific Review Article

    Type
    Blog Post
    Published
    Feb. 16, 2017, 3:30 p.m.
    ...T-cell development, physiology and malignancy." Genes & cancer 6.5-6 (2015): 184. PubMed PMID: 26124919...PMCID: PMC4482241. Additional Resources on the Addgene Blog Writing Scientific Manuscripts: Literature...Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out Our Hot Plasmids Articles to See Compiled...

  22. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...delivery of genetic cargo  By Alyssa NeuhausTo improve delivery and reduce off-target effects of gene therapy...system to improve on-target delivery of genetic cargo Optogenetic proximity labeling and TransitID Tracking...continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, that overcome...10.1038/s41467-023-40788-8. PMID 37612276.   Optogenetic proximity labeling and TransitID By Mike Lacy...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into...43R] has been one of the hottest antibodies in Addgene’s collection since we began distribution and has...[K28/43R] under the Application section of the Addgene antibody page, and learn more about the whole-brain...

  23. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...to make sense of biology, with a special focus on gene editing techniques. References and resources References... versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. https://doi.org/10.1038/s41576...DeepSpCas9, a deep learning–based model with high generalization performance. Science Advances, 5(11), eaax9249...). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https://...j.cell.2023.03.034 Resources More resources on the Addgene blog How to Design Your gRNA for CRISPR Genome ...and Flexibility to CRISPR Editing Resources on Addgene.org CRISPR: PRIME Edit Collection The CRISPR Guide...

  24. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ...my students published their work identifying new genes required for stem and progenitor cell maintenance...incorporated a discovery-based lab on planarian regeneration into one of my own courses, I was excited to...be authentic – students should be attempting to generate new knowledge that will be of genuine interest...Provost’s course recently used a combination of phylogenetic analysis, structural prediction, and bench work...recombinant enzymes in bacteria, purified them to near homogeneity using a 6XHis tag, and compared their specific...

  25. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    Published
    Dec. 13, 2016, 3:30 p.m.
    ...studies show that certain mutations of the BAP1 gene play a role in susceptibility to developing mesothelioma...Additional Resources on the Addgene Blog Find out More About Cancer and Addgene's Cancer Collection Pages...others never do, but we do know that there are some genetic factors involved. For example, multiple studies...research is being done to see what, if any, other genetic factors may be in play. However, there is no question...options: Early detection, immunotherapy, and angiogenesis inhibitors New research is also going into two...bevacizumab (Avastin). This drug (known as an angiogenesis inhibitor) prevents the growth of new blood ...the use of asbestos, thus ensuring that future generations will be protected from this dangerous material...

  26. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...the expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a...PMCID: PMC6061786. Additional resources on the Addgene blog Learn the basics of Golden Gate assembly ...other plasmid cloning techniques Resources on Addgene.org Find basic molecular biology protocols Find ...

  27. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...in the fields of gene and cellular therapies.     Additional Resources on the Addgene Blog Read What ...Technical Support Calls Resources at Addgene Visit the Addgene Careers Page Check Out Our Career Videos...You will also help troubleshoot products, give general experimental guidance, and resolve complaints about...willingness to help others with their problems, a general positive attitude, and an open-minded personality...will do well in a technical support role. What general characteristics do jobs in technical support have...covered by the lab based team as part of their general job duties. Bigger companies have dedicated technical...post. Klaus Wanisch is an Outreach Scientist at Addgene. These days he's doing a lot of science communication...

  28. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...from transient overexpression of protein fusions to gene-edited cells can decrease the number of fluorophores...Fluorescent Protein Should I Use? Resources on Addgene.org Generate Fluorescent Protein Fusions Request Plasmids...in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your... dye ligand (e.g., FlAsH, ReAsH) and a short genetically encoded tetracysteine (Cys4) peptide tag could...TMP-tag) – These widely used systems consist of a genetically encoded enzyme variant tag that reacts specifically...such as paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule...strategies have trade-offs between the size of the genetically encoded tag, the speed and selectivity of the...

  29. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...contain either an ampicillin or kanamycin resistance gene. Every step along the way involves good aseptic ...of tube for each plasmid we have at Addgene. Each sample at Addgene has four layers of location which can...pages! Resources Avaliable on the Addgene Blog Read more Inside Addgene Posts! Learn all about Plasmids ...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the... a microcentrifuge tube. Once a sample enters Addgene’s lab, a series of events is triggered. Each step...fluctuates throughout the year (see graph below). Addgene scientists inspect each sample and make sure they...takes over and prepares the sample for entry into Addgene’s repository. For long-term storage in our facilities...

  30. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ... Ti plasmids in Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs... Resources on Addgene.org Download the Plasmids 101 eBook  Browse plasmids at Addgene Read our Molecular...plasmid partitioning system in the bacteria, but generally it consists of a centromere-like region on the...Kamruzzaman et al., 2017). Additional resources on the Addgene blog Read all of our Plasmids 101 blog posts Learn...

  31. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...likely -/-, 50% are -/+, and 25% are +/+ for your gene of interest. Thus, to find a null clone, you should...edited. For many experiments, the end goal is to generate a clonal cell line, so it’s important to verify... for a needle in a haystack before going on to generate and screen individual clones. If a clonal line...al., NAR). This technique uses the trace file generated in Sanger sequencing and a simple algorithm to...with your sgRNA sequence. The TIDE software will generate a graph representing all insertions and deletions... 4 clones. Did you use a dual guide system to generate your knockout? Then you’re in luck! You don’t ... to visualize an edit. Most dual guide systems generate a deletion large enough that it can be visualized...

  32. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...IPTG. T7 RNAP is then available to transcribe the gene of interest from a T7 promoter on the plasmid. Many...::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein...cytosine within a particular sequence. Browse Addgene's curated list of Bacterial Expression Systems Table...DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) General protein expression BL21 (DE3) pLysS* Chloramphenicol...ompT lon hsdSB (rB- mB-) gal dcm rne131 (DE3) General expression; not recommended for toxic proteins ...lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression  BLR (DE3) Tetracycline RecA-deficient...overexpression? The strains described above should generate sufficient expression levels for most purposes...

  33. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...mediated HDR) enables the tagging of endogenous genes close to native expression levels. However, it is...chimera are encoded by DNA. Since researchers can generate almost any DNA sequence in the way that they like...proteins is relatively straightforward. However, generating a fusion while keeping all of the native properties...challenging. In this blog I discuss strategies to generate fusion proteins and highlight some aspects of ... from Aequorea victoria and its variants are genetically encoded fluorescent probes. One of the limitations...restriction enzymes in the multiple cloning site for generating fusions. For the POI-fluorescent protein fusions...sufficient. For the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both...

  34. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...achieved through, for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside... on Addgene.org Ready-to-use recombinant antibodies at Addgene Addgene's Antibody Guide Addgene's Antibody... combines a marker with a signal. For clarity, Addgene uses 'labeling' to refer to antibodies and 'staining...Find streptavidin plasmids and biotin plasmids at Addgene!     Figure 1: Indirect detection of target...signal. Dump gating When you are working with heterogeneous cell mixes, like bulk lympho-/splenocytes, you...Antibody Plasmid Collection Resources on the Addgene blog Antibodies 101: Reading a Flow Plot Antibodies...

  35. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    Published
    March 8, 2021, 2:41 p.m.
    ...do not naturally express the protein or have had gene expression knocked down or knocked out. If you are...antibodies Resources on Addgene.org Find written and video protocols Visit Addgene's antibodies collection...plasmid encoding your protein of interest from Addgene, transfected it into your target cells, now what...wet, semi-dry, and dry but all follow the same general principle where the gel and membrane are “sandwiched...may even find plasmids encoding antibodies from Addgene that you can make on your own.  To narrow your ... found this blog helpful be sure to check out Addgene’s blog for overviews of other antibody topics. ...10.4103/1947-2714.100998 Additional resources on the Addgene blog Find other tips and tricks for molecular ...

  36. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...which come from naturally-occurring immunoglobulin genes, recombinant antibodies are plasmid-based. Their... What is an Antibody? Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...in the DNA of the hybridoma can occur, known as genetic drift, meaning that over time, the antibody you...monoclonal antibodies, recombinant antibodies are a homogeneous population. However, unlike monoclonal antibodies...antigen binding, the variable regions alone can be generated as a fusion protein. An ScFv is made up of the...detect the presence of a single protein from a heterogeneous mixture - for example, cell lysate or media ...

  37. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ... 18 lung cancer patients. This study used CRISPR gene editing to knockout PD-1 on immune cells harvested... Cell heterogeneity Tumors are composed of a wide variety of cell types. This cell heterogeneity is an...cells that escape the elimination stage acquire genetic alterations and alter cell-surface antigen production... promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates the... immunotherapy. Using activated DCs ex vivo to generate cytotoxic effector T cells, and targeting specific...played a huge role. For instance, researchers are genetically altering tumor cells and immune cells to knockout...element of successful tumor progression. A tumor’s genetic composition is constantly in flux and is dependent...

  38. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...procedure and application to the major ALS/FTD disease gene C9ORF72. eLife, 8, e48363. https://doi.org/10.7554...101: Validation Resources on addgene.org Addgene's Antibody Guide Addgene's Antibody Protocols... outlined below. Knockout cell line approach Genetic knockout-based validation can conclusively prove...you are working with cells that are amenable to genetic knockout, this represents a very robust approach...to start with a screening procedure in easily genetically modified cell types (top). The specific approach...10.1038/s41598-019-55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies...

  39. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ...transition from weak, random binding to precision gene editing requires three major steps: RNA loading,...resources on addgene.org  Addgene's CRISPR Guide Addgene's CRISPR 101 eBook  Resources on the Addgene blog Components...from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state...    Figure 3: Crystal structure of S. pyogenes Cas9 bound to guide RNA (PDB ID 4ZT0), shown opaque...sequence isn’t in place yet.     Figure 4: S. pyogenes Cas9 in the apo state (PDB ID 4CMP, top) compared... residues in the CTD can get cozy with DNA. S. pyogenes Cas9 recognizes the PAM sequence 5’-NGG-3’, where...complementary strand is mismatched.   Figure 5: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID...

  40. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    Published
    Nov. 3, 2015, 3:30 p.m.
    ...of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation." Nature biotechnology (2014). PubMed...to select a tool based on your project and not general opinion. I could spend a lot of time running through... Online tools dominate the software space but generally rely on less powerful algorithms to detect off-targets...tools available mostly work on any computer but generally require working knowledge of the command line ...tools require some kind of data input in order to generate results. While different types of input are more...focus on specific organisms. For this reason I generally recommend that laboratories that have already ...PubMed Central PMCID: PMC4262738. Resources at Addgene Get Tips on How to Design a gRNA Read Our CRISPR...
Showing: 521 - 560 of 920 results