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We narrowed to 1,045 results for: ins;

Showing: 411 - 420 of 1045 results

  1. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...approach, which focuses on validating antibodies against markers of the human blood leukocyte populations...cases, we would recommend a knockdown approach instead. In this approach, you’re looking for an antibody...permanent knockdown, consider using shRNA vectors instead. With both siRNA and shRNA, it can be difficult...induce/suppress the expression of many related proteins, not just your protein of interest, which can ...interest, which can be difficult to find for many proteins. In practice, it was necessary for us to combine...primary blood leukocytes. The file shared also contains information about the labeling pattern of several...on human blood leukocytes, including antibodies against these markers. The 371 CD markers characterized...

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

    Type
    Blog Post
    ...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...swapping of fluorescent proteins and multiplexing is simple when fluorescent proteins with limited spectral...FRET reporters are comprised of two fluorescent proteins with very specific spectral properties that limit... and the orientation of the donor and acceptor proteins, which can be difficult to predict. Slower Dephosphorylation...

  3. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...the ability to generate full libraries of mutant proteins. Researchers from the Whitehead lab developed ... Gibson Assembly: An easy-to-use technique that joins homologous DNA fragments in a single-tube reaction...using Gibson assembly is that the longer the oligo insert used, the more likely it will contain random synthesis... enzymes leads to a buildup of uracil and the insertion of uracil into newly synthesized DNA. This labeled...synthesized DNA strand. Each degenerate primer set contains three consecutive randomized bases (NNN) at a ...that the BbvCI enzymes nick the DNA strand that contains their respective recognition site. This means ... matter? Most of the time no, but there are 2 instances where it does. 1) When the backbone vector has...

  4. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...are only expressed when the targeting vector is inserted downstream of a functional promoter (gene traps...2008). While the biology behind this AAV effect remains unclear, it is likely not a coincidence that the...gene. pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR gene,... CRISPR-based approaches for the generation of knockins via homologous recombination with a homologous...application - the creation of sequence-specific knockins - for which the AAV approach is most well suited... CRISPR-based approaches for the generation of knockins, it is not always ideal. Because of these limitations...devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied separately from...

  5. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...reaction that cleaves a nearby RNA sequence that contains a fluorophore that is quenched when intact and...improves sensitivity of the assay. This reaction also contains SSB, strand displacement DNA polymerase, recombinase...fluorescence visualization. In this technique, however, instead of using lateral flow strips to detect the cleavage...Reverse-transcribed RNA samples are amplified by PCR instead of RPA. Unlike RPA, this technique uses Taq polymerase... which is widely available and less expensive. Instead of using the typical PCR machines, the authors ...depend on collateral cleavage by the enzyme and instead relies on its activity on the target regions thus...COVID-19 using CRISPR diagnostics. https://www.broadinstitute.org/files/publications/special/COVID-19%20detection...

  6. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...where they bind to their targets, most commonly proteins. You can then use fluorescent microscopy to look...visualize their target. These antibodies are generated against a specific antigen and then conjugated with a fluorescent...but is not attached to a fluorescent molecule, instead a secondary antibody that targets the primary is...for each primary antibody could get out of hand. Instead, it is more efficient to generate a handful of ... multiplexing is that you can ask how multiple proteins relate to one another in the same sample. For ...see Im, et al. 2019.  Fixation Good fixation maintains sample morphology with minimal impact to the target...spectral properties match your available imaging instruments and the spectral properties do not overlap with...

  7. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...MTRIA for short), where a fluorescent protein is inserted into one of the loop regions of any G-protein-... Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted GFP into an intracellular...calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance for in vivo use and...indicator using small monomeric NIR fluorescent proteins (miRFP670nano and miRFP720) and the Ca2+-sensing...structure-guided engineering of potassium binding proteins, they developed a family of fluorescent potassium...et al. 2022). By fusing two bright fluorescent proteins with different thermal quenching rates (mNeonGreen...Additional resources on the Addgene blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org...

  8. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...plasmid, into which the GOI is inserted, and a Ti "helper" plasmid, which contains Vir genes necessary for T-DNA...gRNA scaffold sequence and allow researchers to insert targeting oligos with minimal cloning required....Dicot Genome Editing Kit from the Stuttmann lab contains a variety of Agrobacterium-compatible, Cas9 containing... T-DNA processing and insertion into the plant genome. B) The Ti plasmid and helper are transformed into...the Vir genes on the helper plasmid and used to insert the intervening region into the plant genome. Created...- promoters used, precise protein sequences or domains, and methods of delivery - the underlying technique...

  9. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...studies or make a reporter construct. Fluorescent proteins (FPs) are also the foundation of multiple important...biological research. Photoconvertible fluorescent proteins Newly developed photoconvertible FPs like PsMOrange...repeat the process with these new photoconvertible proteins. With Sattarzadeh et al.’s discovery that GFP ...After exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. noticed that GFP ...encoding one of the unexpectedly photoconvertible GFP proteins to make a new transgenic line could avoid problems...as well as a wide variety of other fluorescent proteins for your imaging needs. If you need an FP refresher...

  10. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...Llamas Required - Synthetic Nanobodies Against Membrane Proteins More resources from addgene.org  Plasmids...single-domain antibodies genetically fused to fluorescent proteins and used to visualize target antigens in living... transgenic mice that express camelid variable domains (aka nanomice), which gives researchers the “best...made to generate the nanomouse. (VHH = variable domains of heavy chain only antibodies) (B) Nanomice express...widely adopted. So thank you to the camelids who inspired this versatile family of tools, the applications...
Showing: 411 - 420 of 1045 results