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We narrowed to 537 results for: val

Showing: 81 - 100 of 537 results
  1. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...antibodies validated for IP might not work for a western and vice versa.  In addition to validated applications...become the gold standard in antibody validation. This level of validation ensures that the antibody is staining...measure the protein content and normalize to ensure equivalent loading. Denature the samples to their primary...narrow your search, first pay attention to the validated applications listed on the vendor’s website and...and choose an antibody that has been validated for immunoblotting. Different immunoassays detect proteins...lines other than those used by the vendor for validation could contain unintended cross-reactive proteins...interested in making your own knockout line for validation, please view this tutorial from Stuart Orkin'...
  2. Building Global Connections with the International Mentorship Program USA-EUROPE

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    Blog Post
    ...successes. Although all mentorship relationships are valuable and highly recommended, a mentor with international...Joaquin Lilao (Mentee IMP 2014-15, University of Valencia): “At the beginning, I registered at IMP to have...Elena Grosso (Mentee IMP 2014-15, University of Valencia): “It is very inspiring to be in contact with ...Adrian Mancebo (Mentee IMP 2014-15, University of Valencia): “IMP showed me a wide range of professional ...locations in Spain were enrolled: the University of Valencia, University of Oviedo, the European University...completed follow-up self assessments and program evaluations every four months. All surveys were anonymous...scientists, who did not have contact with the program, evaluated survey results. Participants’ responses to the...
  3. Click Biology: How to Click and Play to Explore Biology

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    Blog Post
    ...reactions, one component bearing an azide reacts covalently with another component bearing an alkyne; the...building-blocks of living cells, rapidly forming covalent bonds to specific partners under cell-friendly...can help with this!). Besides requiring stable covalent reactions, other required parameters of the coupling...function to My Favorite Protein (MFP) through rapid covalent coupling. Created with BioRender.com.   Table...SpyTag003/SpyCatcher003 SpyTag/SpyCatcher Rapid covalent bond formation between a genetically encoded peptide... SnoopTag/SnoopCatcher SpyTag/SpyCatcher Rapid covalent bond formation between a genetically encoded peptide...expansion DogTag/DogCatcher SpyTag/SpyCatcher Rapid covalent bond formation between a genetically encoded peptide...
  4. A Career in Grants: How To Become a Grant Professional

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    Blog Post
    ...professional works for or with an organization to evaluate the organization’s grant readiness, identify grant...coordinating the grant application and submitting it, to evaluating the capacity of an organization to receive and...that grant reports are submitted and project evaluations are completed. To enter and advance the profession...grant management, grant administration, and grant evaluation are available through not-for-profits, universities...disciplines, writing, project management, and program evaluation are highly transferable. Use your experience ... development, project implementation, program evaluation and grant management.  Your niche will change...earning a degree in Grant Writing, Management and Evaluation and/or earning the GPC credential from the Grant...
  5. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...you find validated antibodies? Many companies that produce and distribute antibodies validate them in ...organism. But, as with any antibody-based technique, validation and proper experimental design are essential.... reason is cost effectiveness. Generating and validating primary antibodies is a time consuming and expensive...Example steps include: permeabilization, antigen retrieval, and sectioning. Blocking Block is used to...Try to choose primary antibodies that have been validated in either IF,  IHC, or ICC. The biggest difference...will need to use. (To learn more about antibody validation, this commentary by Uhlen, et al. 2016 is a good... antibodies from different companies, such as Validated Antibody Database and CiteAb. The antibody you...
  6. PCR Applications for Copy Number Variation Assays

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    Blog Post
    ...output number (RQ value) matches the copy number of your sample. In that case, an RQ value of one (or, more...would mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so...also serves as a light technical control - an RQ value of 1.5 means you have either made a pipetting error...error or an incredible discovery. The range of RQ values used to call copy numbers should be as small as...determination of copy number, albeit within a range of RQ values as noted above.   In both applications, results... statistics), and the readout gives an absolute value of DNA concentration. By comparing the number of... number can be determined through concentration values, rather than by comparison to a standard curve,...
  7. Meet an Addgenie: Insights into Grad School, Career Paths, and More

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    Blog Post
    ...She told me, “I think that my experience is more valuable at this point. I don't know where a master’s would...Addgenies features on Instagram! Staying true to your values While going through the co-op application process...process, my advisor encouraged me to keep my values in mind as I applied. Thinking about this made me curious...curious. I wondered what my coworkers value and if that is a big part of why they work at Addgene. I posed...of getting scooped, she said. Reflecting on her values, she decided that this “is not [her] view of science... she stuck with it. Lab Manager Shannon Rinaldi values loving what she does. Her parents told her that...different career paths, successes and failures, values, and more. I have developed connections with my...
  8. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

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    Blog Post
    ...experimental hurdles I would encounter. This was invaluable to me as a bench scientist working at an institution...opportunities weren’t always available. Even more valuable, however, was when she invited me to lunch for...not participate but provided moral support was invaluable.  Mixing metaphors and scientists - Culturing...lineages share genes that can improve chances of survival for an individual, and eventually, a community...community. The experiences of the mentors were very valuable when shared with the next generation of scientists...together with the society to bring an even more valuable program to Microbe 2020 in Chicago.  Finding our...conferences can use to foster a culture that brings more value to ALL their members.  Shoutout to our mentors and...
  9. Antibodies 101: Normalization and Loading Controls for Western Blots

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    Blog Post
    ... controls are equally strong, so the normalized value simply reports which POI band is stronger. The right...in condition 1 than condition 2, the normalized values for the two conditions are equal. Created with ...these other sources of error. It’s important to validate these assumptions for your particular experiment...analogous green “condition 2” points by the exact same value. For the two solid POI points, this difference is...detection range. As a result, they have almost the same value on the band intensity axis. This effect causes misleading...reflect massive differences in protein abundance. Validating the assumptions for different loading controls...assumption is likely true in many cases, you should validate it for your particular setup. We recommend starting...
  10. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

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    Blog Post
    ...This dataset currently includes 1049 community-validated reagents from 20 contributors, including 61 fluorescent...for the performance of a reagent, e.g., antigen retrieval conditions, dye inactivation details, tissue preservation...preservation, target tissue, detergent, antigen retrieval conditions, and more. Users can also recommend...an author.  The IBEX Knowledge-Base is an invaluable resource for anyone performing multiplexed imaging...used IBEX and wants to preserve and share their validation work. Everyone who contributes to the IBEX Knowledge-Base...named on the Zenodo release in keeping with their value of shared ownership. Go check it out! References...
  11. Plasmids 101: Restriction Cloning

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    Blog Post
    ...ligase to covalently link the fragments together at the expense of ATP (see below, covalent bonds are ...restriction enzymes. An enzyme, DNA ligase, then covalently binds the plasmid to the new fragment thereby... to make bonds between the insert and backbone covalent.  Before beginning the restriction digest...orientation. You then add ligase to the mixture to covalently link the backbone and insert and, PRESTO, you...vector + insert plate (unsuccessful 2), or roughly equivalent numbers of colonies on each plate (unsuccessful...colonies on your backbone plate (greater than or equivalent to backbone + insert, unsuccessful 2 and 3 above...
  12. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...of a high throughput survival screen found that either technology was equivalently precise, but each method...same gene. However, recent work has called this prevalent assumption into question. Specifically, CRISPR-generated...DNA template for the RNAi trigger is the most prevalent method due to ease of use and capacity to generate...must be individually engineered and empirically evaluated for their gene silencing efficacy, thereby requiring...engineered a positive selection reporter whereby survival itself is indicative of active gene silencing...., potent gene silencing of APT is required for survival. By coupling silencing of APT and any gene target...
  13. Technical Design of a Western Blot

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    Blog Post
    ...recommend using an antibody validated for your target in your specific assay. If validated antibodies aren't available... different than the validation assays, we recommend performing your own validation assays. If you’re using... the manuscript and supplementary figures for validation data, or follow the references to find the original...original publication and ensure that it includes validation data relevant to your application — and double-check...significantly from lot-to-lot, and have to be re-validated each time a new lot is purchased. Monoclonal antibodies...manufacturer, each rAb in a mixture can be individually validated. ‌rAbs are not as commonly available as monoclonal...
  14. Screening for Successful Genome Editing with Digital PCR

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    Blog Post
    ...genome editing. When it comes time to (hopefully) validate successful mutation of your favourite gene, there...guide you through the rough waters of mutation validation, such as this "CRISPR 101" post. Next-generation...While these methods paved the way for mutation validation, we found these assays frustrating to work with...post, we’ll introduce digital PCR as an emerging validation technology. Digital PCR has several advantages...editing workflow. Digital PCR comparison to other validation methods As mentioned, these assays have several...harnessing the power of digital PCR screening to validate your genome edits, this paper provides detailed...
  15. Antibodies 101: Isotypes

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    Blog Post
    ...means each IgM molecule has a binding capacity or valency of 10 antigens. An extra constant domain replaces... is unknown. It has a monomer structure with a valency of two and has no subclasses. IgG IgG is the most...subsequent time. It is expressed as a monomer with a valency of two. There are four IgG subclasses, numbered...subclasses and they are expressed as monomers with a valency of two.  IgA IgA antibodies are found in blood,... common antibody after IgG; but it's the most prevalent antibody in secretions (e.g. tears, saliva, mucus.... This isotype is most commonly a dimer with a valency of four. There are two IgA subclasses: IgA1 and...
  16. Live and Let Dye: Self-Labeling Protein Tags

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    Blog Post
    ...catalytic activity and instead become trapped in a covalent bond with its substrate. Then, all it takes is...on the human DNA repair protein AGT that gets covalently linked to derivatives of its substrate O6-benzylguanine... 2005). Although this tight interaction is not covalent, later work showed that introducing a Leu28Cys...Leu28Cys mutation in eDHFR does lead to covalent binding of the TMP-tag ligand (Gallagher et al., 2009). HaloTag...Sheetz, M. P., & Cornish, V. W. (2009). An In Vivo Covalent TMP-Tag Based on Proximity-Induced Reactivity.... Adams, S. R., & Tsien, R. Y. (1998). Specific Covalent Labeling of Recombinant Protein Molecules Inside... Johnsson, K. (2003). A general method for the covalent labeling of fusion proteins with small molecules...
  17. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ...nucleic acids, or glycans. Fluorophores can also be covalently attached to self-labeling proteins. Some common... of a genetically encoded enzyme variant that covalently reacts with a small substrate ligand motif attached..., maintaining intensity at ~85% of its original value after 60 minutes of illumination, highlighting its...(Sridharan et al., 2014). It's also crucial to evaluate how the addition of fluorophores affects the functional... or the degree of labeling (DOL), and thorough removal of unconjugated dye molecules are critical for ...filtering settings of the equipment. Systematic evaluation of commercially available dyes with different...Kurumizaka, H., Nozaki, N., & Kimura, H. (2014). Evaluation of chemical fluorescent dyes as a protein conjugation...
  18. Hot Plasmids and Viral Preps - May 2021

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    ...https://doi.org/10.1126/science.abf9302     Validated recombinant antibody toolbox for aminoacyl tRNA... (aaRS) have been created, characterized, and validated by the Gräslund lab. Recent findings suggest that...inflammation and autoimmune diseases, so these validated antibody tools could be used to study the role...slow off-rate binders. The best candidates were validated by IP-MS using HEK293 cell lysate where the antigens...system. Image from Cheah et al., 2021.   After validating this method through monitoring GFP degradation...Dimidschsteind lab provide expression selectively in Parvalbumin containing cortical interneurons. We now provide...
  19. CRISPR 101: Mammalian Expression Systems and Delivery Methods

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    ...non-homologous end joining (NHEJ) or HDR. EGFP validation of sgRNAs: Can evaluate component delivery and sgRNA efficacy...Target DNA reporter system: Can evaluate component delivery with validated tools.  Special thanks to Joel...delivery be more suitable? Determine how you will evaluate the outcome: Will you be detecting insertions/...testing tools: Traffic Light Reporter System: Can evaluate lentiviral component delivery as well as genome...
  20. Antibody Data Repositories and Search Engines

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    ...May include validation data CiteAb Any Any Search engine May include validation data  Biocompare...help: Antibody data repositories, which share validation data, and antibody search engines, which allow... you're looking for an antibody, especially a validated antibody, we recommend using antibody search engines...from many vendors, while data repositories share validation and experimental data to help you decide if the...
Showing: 81 - 100 of 537 results