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We narrowed to 486 results for: mme

Showing: 21 - 40 of 486 results

  1. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ... or a combination of the two. There are many commercially available options to choose from.  Transfer ...staining or other reversible protein staining immediately after your transfer. If you are expecting low...blockers, and the manufacturer will usually include recommended blocking conditions on the spec sheet. If you... background after following manufacturer’s recommendations, you can try either longer blocking times or...antibody information doesn’t contain blocker recommendations, you may want to try different protein blockers...make up a fresh batch.  Antibodies We always recommend either using an antibody validated for your protein... well. Dilutions Most antibodies come with a recommended dilution and/or range (typically between 1:500...

  2. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...five pillars of antibody validation, and it is recommended to screen multiple candidate antibodies when ... interest when expressed at endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly... to produce KO cell lines, some of which are commercially available. Various proteomic and transcriptomic...knockout approaches. In these cases, we would recommend a knockdown approach instead. In this approach...individual experiment.   Figure 3: Our recommendations for an approach that may be used to determine... onto the five pillars, with some specific recommendations for how to use these in practice. This ...received in-kind support to produce this data from commercial antibody manufacturers. References and resources...

  3. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...membrane protein targets in as little as 3 weeks (Zimmermann et al., 2020). This selection platform was designed...interest using the Sybody Generation Toolbox and commercially available reagents in three major steps - all...can be randomized for creating the libraries (Zimmermann et al., 2018). Find the Sybody Generation Toolbox...sybodies against membrane proteins. Image from Zimmermann et al., 2018. Ribosome display The Seeger ...members). This selection step involves using a commercial kit for in-vitro translation of the mRNA libraries...identifying unspecific and low-affinity sybodies (Zimmermann et al., 2020). This step of the protocol starts... . https://doi.org/10.1016/j.cell.2020.04.031 Zimmermann I, Egloff P, Hutter CAJ, Kuhn BT, Bräuer P, Newstead...

  4. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    Published
    Aug. 8, 2019, 1:19 p.m.
    ...to address the comments to “you” rather than “the authors.” By addressing the comments to “you” I can ... figures). After finishing my section-specific comments, I re-read what I wrote about the paper after ...in the first (and clunky) version of my general comments and adjust them if my opinion changed after this...that there is another human who is reading your comments. Receiving a peer-review is already a humbling...easily check if I would have formulated specific comment differently if I would be speaking to a colleague...young scientists that are reading your peer-review comments will learn from them and propagate this culture...

  5. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ... to lyse the cells to let the virions out. We recommend using a Tris-based lysis buffer and sonication...Grimm et al., 1999), quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet digital PCR (ddPCR) (Lock...freeze/thaw cycles (Lock et al., 2010). It is recommended that you measure your infectious titer, since...standard labs. When determining titers by qPCR, we recommend using primers targeting the viral genome or inverted... quantitation blog post.  We generally do not recommend using ELISA-based titering assays, since those...August 2023 References and resources  References  Aurnhammer, C., Haase, M., Muether, N., Hausl, M., Rauschhuber...10.1089/hum.2010.107 Zen, Z., Espinoza, Y., Bleu, T., Sommer, J. M., & Wright, J. F. (2004). Infectious titer...

  6. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...depositor recommendations to grow a plasmid at 30℃ or room temperature. These recommendations also often...strain. For most applications, Addgene uses and recommends DH5𝝰! This strain consistently gives good, high...degradation of your prep. Many plasmid prep kits recommend performing an extra wash step to remove endonucleases...common lab E. coli strains on the blog, and also recommend this handy guide from OpenWetWare for a more extensive...plasmid prepping. Be sure to stay within the recommended range of the filter or you might inadvertently...cultured in media containing half the normally recommended antibiotic concentration. Add supplements to ...

  7. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    Published
    July 14, 2017, 2:30 p.m.
    ...Lader Commercial DNA Ladder Penn State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid... 1 kb DNA ladders, respectively. Unlike many commercially available ladders, the 100 bp ladder works well... $20-30 Cost Comparison: While convenient, commercial ladders cost between $200-$500 for 1000 lanes...worth of the 100 bp or the 1 kb ladder. Using a commercial plasmid prep kit increases the costs to $20-30... per ladder: still significantly cheaper than commercial ladders. Are you interested in making your own...

  8. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...your work, downloads, commercialization of patents and letters of recommendation. EB-1A extraordinary ... EB-1B cases, Casseus Law recommends 6-8 letters.  For these recommendations, Casseus Law suggests a mix... pressure off having to find the perfect job immediately after you complete your Ph.D. or postdoc. Foreign.... High Salary –not relevant to most postdocs. Commercial successes in the performing arts- not relevant...an expert is someone who has cited your work. Immediate steps that you can take to prepare for the green...application process. To answer specific questions, we recommend that you reach out to experienced legal counsel...

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

    Type
    Blog Post
    Published
    May 25, 2021, 2:29 p.m.
    ...James Trimmer of the University of California Davis School of Medicine on this grant. Dr. Trimmer has created... ready-to-use-proteins to facilitate use. Dr. Trimmer’s lab will ensure the quality of the antibodies ...Addgene to bring this resource to life,” says Dr. Trimmer. But in order to create such a resource, we need...Initiative has been funding laboratories, such as Dr. Trimmer’s lab, to create recombinant antibodies for use ..., Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P, Frenzel A...

  10. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    Published
    June 2, 2016, 2:30 p.m.
    ...code remains in tact! Take notes Comment in your scripts: Comments on what a given line of code does ...the code. # This is a comment in python, R, perl, and ruby // This is a comment in C++ and Java Command...invaluable. In the top directory of every experiment, immediately write a text document with a brief description...and error, and by asking for tips from others. Comment below to add your tips! Many thanks to our guest...

  11. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ... and Morrison is nearly identical to that of commercially available kits but instead of binding DNA to...yield and quality similar to that obtained with commercial kits, but in less time. The authors estimate ...completed in 20 to 30 minutes. Additionally, while commercial plasmid prep kits come in discreet sizes (mini...cheaper, but also faster and more flexible than commercial kits.  DNA gel extraction with glass milk Starting...where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based kit, this cellulose-based... mentioned in today’s post? Let us know in the comments! Additional resources on the Addgene blog Read...

  12. Transferable Skills Guide: Time Management

    Type
    Blog Post
    Published
    Dec. 5, 2017, 1:52 p.m.
    ... you might need to do. In most cases, I would recommend the second option for the experiment above, but...cells later. Eating with friends has a somewhat immediate deadline - even the best of friends get hangry... everything in a Word or google doc that I can comment and make notes on as I progress. I’ve also recently...are far too distracting. (As a side note, I’d recommend the book, “Bored and Brilliant” for learning more...Harvard Business Review article on procrastination recommends testing out how much time you can possibly handle...suggestions, please be sure to note them in the comments! Additional Resources on the Addgene Blog Management...

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

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...uploaded to Addgene. We also recommend inserting self-cleaving hammerhead ribozymes directly upstream ...construct your own two-input or three-input GRSMs, we recommend using overlap extension PCR and Gibson assembly...reliability of the parts used to build them. We recommend using strong, directional terminators and promoters...transcribed gene and its 5’ UTR. Various self-cleaving hammerhead ribozyme parts are described in Nielsen et al...implement the same gene regulation program. We recommend testing multiple register designs for the same...

  14. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

    Type
    Blog Post
    Published
    June 8, 2023, 1:15 p.m.
    ...share both their successes (recommended reagents) and failures (not recommended reagents) along with information... conditions, and more. Users can also recommend or not recommend a particular reagent in the given conditions...documentation that agrees or disagrees with the recommendation. The goal is to easily share and find what ...

  15. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...brain barrier we would currently recommend PHP.V1 or AAV-BR1. We recommend you keep an eye out for emerging... to avoid "leaking" into the brain, would you recommend using the PHP.S capsid over the PHP.eB (the capsid...capsid we have been using)?Would you recommend any liver specific capsid, particularly one that can target...in the blood-brain barrier?   A: For liver, we recommend using one of the natural serotypes such as AAV9...

  16. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    Published
    Oct. 25, 2016, 2:30 p.m.
    ...institutional arguments were not as convincing as the recommendations for postdocs to be on higher salaries (which...Dame sets no minimum, stating that while they recommend NRSA levels, postdocs must be at least at the ... possible outcome: that many postdocs will immediately lose jobs; that tenured professors/mid-career...system sustainable. For years, there have been recommendations that postdoc salaries be raised to $50,000 ...There was ample opportunity to accept these recommendations, and make gradual adjustments towards this ...

  17. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    Published
    Oct. 20, 2016, 2:30 p.m.
    ...questions and comments go up early in the day, and the moderators were on top of any comments that were at...Anything”) on r/Science. You can see some of Joanne’s comments on the AMA process below, but we also wanted to... scientists who will find it useful, I highly recommend doing an AMA! Before I turn it over to Joanne,...resources. I’ve said this before, but I also highly recommend taking every opportunity you get to speak publically...

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

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...protein expression and some of the most popular commercial strains designed to avoid them. Why do I need...gal dcm rne131 (DE3) General expression; not recommended for toxic proteins BL21-A1 Tetracycline Arabinose-inducible...expression plasmid from these strains is not recommended as these auxillary plasmids may be isolated during...interest from a T7 promoter on the plasmid. Many commercial strains carry the DE3 lysogen, as indicated by...favorite protein expression tips and tricks! Leave a comment below or browse Addgene’s collection of empty backbones...

  19. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...Pre and Post (Post recommended) UV Light 0.1 GelGreen Pre and Post (Post recommended) UV or Green (~500nm...important to digest plenty of starting material. We recommend 1.5-2μg of insert and 1μg of plasmid backbone....space to cut out the bands. Because of this we recommend that you use a wide gel comb, run the gel on the...covalent bonds are indicated in red).     We recommend around 100ng of total DNA in a standard ligation...

  20. A Scientist's Guide to Ebola

    Type
    Blog Post
    Published
    Oct. 16, 2014, 3:01 p.m.
    ...antibodies and inhibitors of attachment and fusion. Recommended Reading For those looking to delve deeper or ...there are plenty of online resources available. I recommend a few: -The recently established Ebola Genome ...EBOV’s past, present, and future, don’t miss Carl Zimmer’s coverage on the subject in the New York Times ...resource to add, please tell us about it in the comments. ...
Showing: 21 - 40 of 486 results