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Showing: 361 - 390 of 846 results

  1. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ...a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such as horseradish...level of antigen present in the sample and is calculated by extrapolating from the standard curve. Because...include undesirable molecules that may bind non-specifically to assay components and increase background ...amount of antigen present in the sample and is calculated by extrapolating from the standard curve. Scientists...antibodies are used.  Competitive ELISAs Scientists typically use a competitive or inhibition ELISA when measuring...While the precise setup of the method can vary, typically users first coat the multiwell plate with a reference...

  2. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...DnaA binds multiple specific sites within the ori called DnaA boxes. These boxes come in both high- and ...Created with BioRender.com. Once DnaA breaks the local hydrogen bonds holding the DNA together, it recruits...itself. RepA binds to directly repeated sequences called iterons located near the ori and directly interacts... the protein to be translated. An antisense RNA called Inc, also encoded by the ColIb-P9 plasmid, is complementary...between iterons on different plasmids in a process called "handcuffing," which blocks replication. ColIb-...Replication Browse other Plasmids 101 Posts Get practical molecular biology help with our Molecular Biology...

  3. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    Published
    Sept. 5, 2017, 1:58 p.m.
    ...combining these two strategies allows you to specifically incorporate homo- or heterozygous mutations....in the field at the time but had never been systematically analyzed: introducing silent CRISPR/Cas9 blocking...DNA region? To this end we developed a technology called CORRECT, which, by undergoing two rounds of gene-editing...efficiently incorporated during HDR. We decided to systematically characterize this phenomenon and found a general...two successful mutations in a single cell. We calculated the likelihood of obtaining homo- and heterozygous...University. He currently works to understand pathological mechanisms of Alzheimer’s disease by developing...

  4. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...cryogenic electron microscopy (cryo-EM) protocols typically require high concentration and purity of the target...limitations, the Funabiki Lab developed a technique called Magnetic Isolation and Concentration (MagIC)-cryo-EM...circles), magnetic bead (*), and spacers are labeled; scale bar: 100 nm. D) 3D model of the obtained structure...right). B) Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of ...orientations, this could be overcome with methods such as chemical crosslinking or tilted-stage imaging. A truncated...evolved recombinases and prime editing. Nature Biomedical Engineering, 10.1038/s41551-024-01227-1. doi:...

  5. Addgene Stands with the Scientific Community

    Type
    Blog Post
    Published
    April 23, 2025, 1:30 p.m.
    ...transformative biological discoveries and life-saving medical breakthroughs and undermine critical research ...Addgene as a trusted partner for archiving their biological materials during funding transitions. Enabling...private partners to create additional pathways for critical scientific work to continue.  Moving Forward ...

  6. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...one be interested in state machine technology? Biological research has produced a massive amount of information...processes that are influenced by temporally ordered biochemical events. How do RSMs work? The state of an RSM...prescribed region of DNA termed the “register”. Each chemical input to an RSM drives expression of a distinct...to record all permuted substrings of a set of chemicals. Specific examples of arrangements that should... record all permuted substrings of up to seven chemical inputs are given in Table S2 of our paper. The... He is broadly interested in biocomputing and biological circuits.     References 1. N. Roquet et al.,...

  7. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...-orbital AAV injection at 4 weeks. Cortical layers (L1-6) and scale bar are indicated. B) Specificity,... core, 2–4 glycosaminoglycans (GAG), and are (typically) tethered to the cell surface by a GPI-anchor.... types in the brain can be a challenging — but critical — factor for successful experiments or therapies...enhancer-AAV vectors targeting specific classes of cortical and striatal interneurons (Furlanis, Dai, Leyva...PV-positive cells out of all BiPVe3-dTom cells. C) Confocal stack of a biocytin-filled cell expressing BiPVe3...

  8. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    Published
    July 26, 2018, 12:46 p.m.
    ...Translocation Reporters (KTRs) whose cellular localization serves as a proxy measurement of kinase activity...KTRs) Kinase Translocation Reporters (KTRs) are genetically encoded fluorescent kinase reporters. A KTR is...which is further fused to a bipartite nuclear localization signal (bNLS) and a nuclear export signal (NES...phosphorylatable. Prior to phosphorylation, the KTR localizes to the nucleus, but following phosphorylation ...breakdown (without the nucleus, there’s no nuclear localization to monitor). Whether you’re looking for a ready-to-use...

  9. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 7, 2017, 1:58 p.m.
    ... first, which he called aequorin, gives off a faint blue light when it binds calcium ions, and the second...second, which we now call GFP, absorbs that blue light and glows green. In the late 1980s, another researcher...gene transcription, as well as to track protein localization. And as it turns out, he was right! He began.... He even obtained samples of certain ones from local pet shop aquariums in Moscow where he lived! His...

  10. Editor's Choice, September 2016

    Type
    Blog Post
    Published
    Oct. 7, 2016, 4 p.m.
    ...re tired of calculating dilutions or you can’t quite remember the hazards of that chemical in your obscure...surpassed 60,000 views for the month of September! Historically we do better in September than in the summer..., Lianna Swanson, and researcher at Sanofi Pharmaceuticals. This post highlights a rare disease, Fibrodysplasia...can find the rest of our September posts in chronological order here: Four Things I Learned at the 10...

  11. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...human speech near the laminar airflow, the human scalp, sneezing, coughing, etc. Also, a constant in-flow...contamination Detection of mycoplasma by naked eyes or optical microscope can be very difficult, so how do you...cells detect mycoplasmas, they trigger a series of chemical reactions and this induces a noticeable color ...contamination, before using the cell lines for assay or analytical purposes. Use 0.1-micron filters as are they ...Fumigate the laminar airflow and the lab facility periodically. The CO2 incubators, used for maintaining cell...cleaned regularly and checked for contamination periodically. For more tips, check out Addgene's video Getting...

  12. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ...depositors; Gizela, the lab member who knows how the physical kits are made and shipped; Chiara, the tech transfer...repository, performing quality control, and providing technical customer service. Team members are highly trained...problem? Test a hypothesis? Whether you officially call it a “project team” or not, most scientific collaborations...But this is a blog post, so here are just a few critical features based on my own experience: Leadership...should be on the team. As a team leader, I’ve (diplomatically) booted people off teams. I’ve also kicked ...like meetings, but run well, meetings can be a critical part of any successful team. For reasons I don...

  13. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...by the use of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD...these complexes are so robustly expressed that anatomical assessments can be easily performed too. While...identified this feature of AAVs as a new tool for neuroanatomical tracing (11). With the ability to produce neuron...Retrograde AAV Preps at Addgene 2) Comparison of genetically engineered receptors (i.e. chemogenetics vs. ...should strengthen its appeal for the scientific and medical communities. Many thanks to our guest blogger ... Floris G., et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring...

  14. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    Published
    Jan. 21, 2020, 2:30 p.m.
    ...published data by biomedical researchers and ready access to stably archived large-scale datasets by computational...new freely available resource developed by the Biological General Repository for Interaction Datasets (...CRISPR screen results available to the entire biomedical research community, but also allows cross-interrogation...relevant to screen result interpretation. The analytical method for score assignment and significance ...

  15. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ... developed an improved voltage indicator (GEVI) called JEDI-2P. Using a custom, high-throughput screening... the protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is ...approach and outcomes of the screen for improved genetically-encoded voltage indicators (GEVIs) based on the...optogenetic silencer in experiments with mouse cortical neurons, far out-performing previous attempts ...selective for K+ over other Na+ and other cations. B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer...

  16. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...The biomedical field is often concerned with understanding the cause of diseases and how to treat those...optimization is performed to take the drug to the pre-clinical and human trial phases.   So how does CRISPR factor...enables researchers to select and generate physiologically relevant models. In complex diseases, it can...standard is to test the drug on models which are genetically proficient and deficient for the target. CRISPR...discovery – it’s an exciting time to be in the biomedical research field! References and Resources References...

  17. Antibodies 101: Epitope Availability

    Type
    Blog Post
    Published
    Jan. 11, 2022, 2 p.m.
    ...antibodies in your lab. Perhaps you’re even in that mythical place where antibodies are well-organized, documented... sample prep. This involves using heat and/or chemicals to break some of the bonds keeping the protein...epitope (right). Created withBioRender.com.   Biological Processes  If you’re feeling relieved that you...the available epitopes could be affected by, or critical to, the assay.   Next, look for antibodies that... also working for a Western because they both typically use denatured protein samples. If, however, you...

  18. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    Published
    May 25, 2021, 2:29 p.m.
    ...reproducibility and an inability to improve upon these critical research tools. Addgene is excited to announce...reagent resource for the neuroscience community called NABOR (Neuroscience AntiBody Open Resource). We...partnered with Dr. James Trimmer of the University of California Davis School of Medicine on this grant. Dr. Trimmer...in neuroscience research today, but there is a critical unmet need for an open-access, molecularly defined...such as plasmids and viral vectors. However, it typically takes a few years for a new project to reach a...

  19. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    Published
    Aug. 18, 2023, 4:15 p.m.
    ...also have a lipid bilayer surrounding the capsid, called an envelope. Viruses without an envelope are referred...non-enveloped, while viruses with an envelope are called enveloped viruses (Figure 1). Like bacteria, only...individual viral particles. A single viral particle is called a virion. Finally, viral vectors, which are used...labs. Viral vectors are viruses that have been genetically modified to either limit or completely eliminate...Viral Applications Viral Vectors 101: Viruses as Biological Tools Viral Vectors 101: Viral Vector Elements...

  20. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    Published
    April 25, 2023, 1:15 p.m.
    ...constant regions are identical for antibodies within certain classification groups called isotypes.  Why does...that of a different group (Fig. 1). This process, called isotope conversion, gives users the flexibility...antibodies are plasmid-based and are created synthetically or derived from hybridoma antibody sequences...antibodies affects binding properties. In Journal of Biological Chemistry (Vol. 269, Issue 17, pp. 13048–13055...

  21. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    Published
    Oct. 29, 2015, 2:30 p.m.
    ...hereditary determinant”. His proposal, however, was basically ignored. A separate term, “episome”, defined as... plasmids have greatly impacted many areas of biological research and have been key in advancing our knowledge...Joshua. "Cell Genetics and Hereditary Symbiosis" Physiological Reviews 32.4 (1952) 403-430. Pubmed PMID: 13003535...PMC2134509. Cohen, Stanley N. et al. “Construction of Biologically Functional Bacterial Plasmids In Vitro.” Proceedings...Nomenclature for Bacterial Plasmids: A Proposal.”Bacteriological Reviews 40.1 (1976): 168–189. PubMed PMID: ...

  22. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    Published
    Aug. 25, 2015, 2:30 p.m.
    ...Harvard Medical School and runs everything science (CSO). Clinton is a cost analyst at Dow Chemical (a Fortune...a company in the health sciences without those critical three letters after your name (P-H-D).  Finally...two projects, including one led by a talented biomedical engineering team from MIT.   I would like to ...connections and finding mentors -- well, it's time you call upon these people for guidance.  Before even beginning...

  23. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...has a maximum of one gene knocked out. In this genetically heterogeneous population, cells behaving differently...knockout mice for (all of the genes involved as I recall). The lab then used these knockout mice to show...cells we study are migratory, and their exact localization is important for their function. Their migration... you could majorly accelerate progression into clinical trials. Secondary screens  If you have generated...genes you’d like to test for phenotypic effects. Typically the phenotype would be studied in vitro, but there...The different labs use different algorithms to calculate which would be the best gRNAs. Interestingly, ...

  24. Networking Follow-up: I Met a Person. Now What?

    Type
    Blog Post
    Published
    Jan. 12, 2021, 2:15 p.m.
    ...understood what I was going through. (Parenthetically, when I called the lab to ask my benchmate to take...within a few days or a week) so they can easily recall the encounter. As a side note, my guess is that...; focus on the nice people who do respond. Don’t call or email multiple times if you don’t get a response...years ago using a now defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is...

  25. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    Published
    July 21, 2020, 1:15 p.m.
    ...might not have to do the calculation yourself as many fluorometers will calculate your sample concentration...than 1.5. Then, using the A260 reading, you can calculate the DNA concentration. Generally, A260 of 1.0 ... start by pouring your gel containing a DNA intercalating dye (ex: ethidium bromide) and choosing a DNA...quantified over time using a fluorescent dye that intercalates into the DNA such that smaller fragments are...

  26. Scientists Map the SARS-CoV-2-Human Interaction Network

    Type
    Blog Post
    Published
    April 9, 2020, 1:15 p.m.
    ...these proteins are FDA-approved or in clinical trials or preclinical studies already. We are following up...expand to other cell types, including more physiologically-relevant systems, to catch all the important... Nsp3 and Nsp16, and includes an additional catalytically dead version of the viral protease, Nsp5. Our...Melanie Brewer (right) from the University of California, San Francisco. Manon Eckhardt and Melanie Brewer...

  27. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    Published
    Sept. 28, 2016, 2:35 p.m.
    ...four proteins Oct4, Sox2, Klf4, and cMyc (the so called OSKM factors). As the Yamanaka lab's primary Addgene...field has exploded so quickly, citing the first clinical trial with iPSCs with his collaborator Takahashi...picture perspective: Dr. Yamanaka talked about his clinical trial patients with advanced macular degeneration...Eggan presented work that is leading to promising clinical trials for ALS.   I have an uncle with Parkinson's...the first stem cell line that could be used in clinical trials. The quality control process alone takes...

  28. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...both perform an optimization-scale infection and at least one screening-scale experiment achieving the gRNA...Chavez, researcher at the Wyss Institute for Biologically Inspired Engineering and member of the Addgene... studying. You need to make sure your assay is scalable and robust. What you don't need to do is spend...particular, the Church lab at the Wyss Institute for Biologically Inspired Engineering and the Zhang and Doench...

  29. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    Published
    Nov. 20, 2018, 8:17 p.m.
    ...to identify and reference documents, files, or physical reagents. What are persistent identifiers? Perhaps...persistent and exist online indefinitely. PIDs historically referred to objects that exist online, but this...approach has been adapted for the identification of physical materials that we use in our labs, including plasmids...Bourges for editing this post and sharing her technical expertise in persistent identifiers. Additional...Identification Initiative Compact identifiers for biomedical data How Addgene works to enhance scientific ...

  30. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...several treatments are currently being tested in clinical trials. It is speeding the process of the discovery...saw these two papers,” said the University of California Berkeley’s Jennifer Doudna. “They are the first...continue to be a primary preoccupation of her California lab and many others. “Those off-target effects...cleavage sites are repaired.” Those insights will be critical for scientists’ ability to influence editing efficiency...Application in Genetic Diseases: A Review.” Current pharmaceutical biotechnology. https://doi.org/10.2174/1389201021666200621161610...
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