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We narrowed to 560 results for: mme

Showing: 61 - 80 of 560 results

  1. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...validation of some of these tools created by the Trimmer Lab that will be the first antibodies distributed...constructing the plasmids for expression of mAbs, the Trimmer lab isolated RNA from cryopreserved hybridomas,...can be a threat to future reproducibility. The Trimmer Lab found that in addition to recovering RNA from...lab’s entire hybridoma collection. Instead, the Trimmer Lab moved to direct functional validation by immunofluorescence-immunocytochemistry...subsequently cloned into the expression constructs. The Trimmer lab found that the recognition sequence for the...webpage. Importantly, the methods developed by the Trimmer Lab for construction and validation of R-mAb expressing...with different mAbs in the same experiment. The Trimmer Lab found that they could modify the CH encoding...

  2. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ...cases”. Cases present specific operational or commercial challenges or questions that clients face imminently... and help resolve it through well-informed recommendations. Healthcare consulting firms that specialize... health and economic benefits of novel drugs. Commercial Affairs teams make up a large and diverse group...group of professionals running the marketing, commercialization, collaborative, and financial functions associated...associated with the business. Commercial teams routinely work with consulting firms on projects such as... conversations with R&D, Medical Affairs, and Commercial teams, reflecting the need for all parts of a...crucial for the success of a drug’s clinical and commercial future, and yet are incredibly difficult to find...

  3. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...version of the plasmid (Summers & Sherratt, 1984; Summers et al., 1993; Summers, 1998). Multimeric plasmids... replicate at a higher frequency (Summers & Sherratt, 1984; Summers et al., 1993). These larger plasmids...monomers and various forms of multimers (Summers & Sharratt, 1984; Summers et al., 1993). In these cases, screening...strategies for resolving multimers? Let us know in the comments below or contact us at [email protected]. Resources...electrophoresis. BioProcess Int. 2006 Sep;4(8):38-40. Summers D. Timing, self-control and a sense of direction....1046/j.1365-2958.1998.01012.x. PMID: 9767582. Summers DK, Beton CW, Withers HL. Multicopy plasmid instability...10.1111/j.1365-2958.1993.tb01648.x. PMID: 8361350. Summers DK, Sherratt DJ. Multimerization of high copy number...

  4. Technical Design of a Western Blot

    Type
    Blog Post
    ...Many antibodies will have a recommended blocking buffer and a recommended concentration. That makes the...particularly your primary antibodies. We always recommend using an antibody validated for your target in... are different than the validation assays, we recommend performing your own validation assays. If you’...There are many different lysis buffers available commercially, and most common buffers have recipes available...to preserve specific protein modifications, we recommend doing a little more research into gel chemistry...separation of proteins of different sizes.  Table 2: Recommended gel percentages for various protein sizes ...binding and washes. It’s less sturdy and not recommended for stripping and reprobing, unless you buy specialty...

  5. Finding Your Perfect Job After University

    Type
    Blog Post
    ... is my experience looking for the perfect job. Summer work experience in cancer research After graduating...would be very competitive, and so I applied for a summer role as a Research Scientist at the Institute of...to gain relevant experience. I applied for this summer position as I felt I needed more practical lab ...study of e2Fb, which is a tumour suppressor. This summer role was incredibly valuable because it was a set... need to explain why I changed jobs. I highly recommend taking on a similar position in your field of ...and enjoyed them, so I applied for a role at a commercial virology testing company. This particular company...companies. It was really exciting to work in a larger commercial company; the pay was good and there was a variety...

  6. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... 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...

  7. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...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...

  8. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  10. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ... 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...

  11. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...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 ...

  12. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  15. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ...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...

  16. DNA Purification Without a Kit

    Type
    Blog Post
    ... 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...

  17. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ... 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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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 ...

  20. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...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...
Showing: 61 - 80 of 560 results