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We narrowed to 356 results for: SAM activators

Showing: 141 - 160 of 356 results

  1. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ...be developed and optimized for the samples, antibodies, activation, and assay readout used. You’ll note...antibody. Perform a second series of wash steps. Activate the signaling molecule and perform the assay readout...approach is as follows: Prepare the sample. Incubate the sample with the primary (unconjugated) antibody...if all the primary antibodies are the same isotype from the same species, only one secondary antibody ...remove any unbound or excess antibody. Incubate the sample again with a conjugated secondary antibody, which...primary antibodies indiscriminately, giving them the same output signal. In these cases, it is best to use...not allow for absolute quantitative analysis of a sample. Instead, relative quantification can be done using...

  2. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...interactions can be discovered by fluorescence-activated cell sorting (FACS). The Mode #2 phosphosite library... a program of OPTN/NDP52 recruitment and TBK1 activation to promote mitophagy . Molecular Cell , 60 (1...Szeligowski, R. V., Rogulina, S., Gerstein, M., Sampson, J. R., Isaacs, F. J., & Rinehart, J. (2018). Encoding...

  3. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    ...Isaacman-Beck et al., 2019. Sparse Predictive Activity through Recombinase Competition (SPARC), a genetic...system to use other experimental transcription activation systems, such as LexA/p65 or mCD8/GFP, to allow...allows for the labeling of a subset of cells with the same genetic makeup, overcomes all of these labor-intensive...upstream of the second attP site to cleave and inactivate any read-through transcripts of the effector ...

  4. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ...changes in autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this ...mRFP-GFP-LC3 functions as an autophagy bionsensor via the same mechanism as Rosella: GFP is quenched in acidic ...

  5. Plasmids 101: Cre-lox

    Type
    Blog Post
    ... – placing Cre downstream of promoters that are active only in certain cell or tissue types, during certain... used for a wide range of purposes, including activating an oncogene only in a particular organ, or bypassing...two loxP sites, which can be located either on the same or on separate pieces of DNA. Both 13bp repeat sequences... sites: Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination... reversed.   Deletion: If the sites face in the same direction, the sequence between the loxP sites is...sites undergo recombination with other sites of the same type, but are not cross-compatible. Using different...

  6. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...substantial size of GFP may interfere with the activity or the localization of the fusion protein. Also... visualization of heterotrimeric G protein Gq activation in living cells." BMC biology 9.1 (2011): 32....al. "A FRET-based biosensor for measuring Gα13 activation in single cells." PloS one 13.3 (2018): e0193705... proteins are also a better choice for thicker samples. Another solution to avoid autofluorescence is ...reaction and hence, there is no need to excite the sample. Although bioluminescent probes are usually dim...

  7. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...of the optogenetic regulator leads to spurious activation of the FRET sensor. Researchers have sought to...Here the bioluminescent donor is unaffected by activation of other fluorescent probes but can still transfer...as the excitation wavelengths needed would also activate the optogenetic accuators. The researchers took...NLuc requires 100-fold less protein to produce the same amount of light from an Fluc or RLuc reaction. In...protein that could be co-imaged with GFP using the same wavelength for excitation. The result was the cyan-excitable... K., et al. (5) imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs...and colleagues (8) were interested in using photoactivatable, optogenetic accuators to depolarize or hyperpolarize...

  8. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ... This repeat region associates with the trans-activating crRNA (tracrRNA) which interacts with the Cas9...Modifications of CRISPR RNAs to improve Gene-Editing Activity and Specificity. JACS 144, 12584-12594 (2022).... when scientists talk about their precious RNA samples. RNA is inherently less stable than most macromolecules...CRISPR-Cas9 experiments – the “gRNA” – suffers from the same issues as your experimental RNA. To solve this problem...Another stabilizing sugar modification, found at the same site, is 2’-fluoro (2’-F), which substitutes a fluorine...protein and can increase on-target efficiency. In the same vein, locked nucleic acids or bridged nucleic acids...modification have overcome these woes with photoactivatable and photocleavable gRNAs. Photocleavable guides...

  9. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...variation in color depends on how transcriptionally active the genomic locus of integration is. The white ...TH17 cell differentiation by antagonizing RORγt activity. Nature. 472, 486–90 (2011). PubMed PMID: 21441909...recombination so that the transgenes can be inherited on the same chromosome. Targeted Insertion allows for direct...variants of YFG because the transgenes are found in the same locus across fly lines (i.e. there won’t be positioning...

  11. Distribution to Industry

    Type
    Collection
    ...Mikel Garcia-Marcos 10 plasmids for detecting GPCR activity in a variety of cell types. Protein Purification...partner labs. Learn More ID Recombinant Antibody Reactivity Recommended Applications PI Don't See What You're...allowed to use for distribution. Please see the Sample Agreements on our Technology Transfer Information...

  12. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...fluorescent proteins (FP) require external light activation, you can’t use fluorescence to monitor processes...light used for excitation can cause unintended activation of the optogenetic system. Biosensors based on...external light source. Therefore, especially thick samples may not be appropriate for use with Nano-lantern...Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check Out Fluorescent...

  13. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...hybridomas, immortalized cells created by fusing an activated B cell that produces the antibody of interest ...heavy and light chain, each from a constitutively active CMV promoter. Constructs were then verified by ...detect multiple targets with different mAbs in the same experiment. The Trimmer Lab found that they could...

  14. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...structure can readily fold into transcriptionally active double-stranded form through intra-molecular annealing...expression patterns of AAV8 vectors with broadly active promoters from subretinal injections of neonatal...episomal and does not divide and replicate in the same way the host DNA does. Because of this, fast dividing...McCarty, D. M., Paul E. Monahan, and Richard J. Samulski. "Self-complementary recombinant adeno-associated...

  15. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...recombinant adeno-associated viruses for Cre activated and inactivated transgene expression in neurons.  PubMed...ability to conditionally turn off one gene and activate another. In the field of mouse genetics, FLEx ...between the sites. If a DNA sequence is floxed in the same orientation, the SSR will excise the DNA fragment...reporter with one loxP site and one lox511 site in the same orientation, and placing the additional loxP and...

  16. CRISPR Library Amplification

    Type
    Protocol
    ...use of high voltages, please use caution when activating pulse and follow all specifications described...gel extraction of either the original sample or the amplified sample, followed by reamplification of the...tens to millions of different plasmids in a single sample. They are often used for screening, barcoding, ...appear to contain sufficient DNA for use, pool samples, and continue with analysis. “Sufficient” DNA will...

  17. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...scientists and non-scientists alike, need to stay active and involved if we wish to make positive change...chair of the postdoctoral engagement committee and active member of the university's Postdoctoral Association...this year I had never taken part in any public activism. One of my favorite signs from the march read ...Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369°W – Mark...campus speakers thoughtfully reminded us to use the same critical lens on our scientific community as we ...well-publicized, we might encounter hostile anti-science activists. However, I must say that I was supremely proud... nor would you mention my small error and at the same time pass over in silence his very large one” (http...

  18. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ... the known pathway that were essential for the activity of different ligands. Such knowledge could be ... to other nucleases as well as inhibitory and activating approaches.  Many thanks to our guest blogger...G., et al. "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9...validation screen using selected gRNAs extracted from the same or another library. These different approaches could...Listed you can also easily extract gRNAs for the same gene from several libraries, and identify those ...

  19. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...subsequently destroyed by Argonaute-catalyzed nuclease activity or their translation disrupted through physical...the appropriate plasmid and dsRNA expression is activated through the  induction of T7 polymerase encoded...Students can produce DNA templates via PCR off of the same plasmid and use these to create riboprobes for in...WISH protocol involves preparation of the tissue sample, several washes, and temperature changes to visualize...

  20. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...could in theory promote off-target binding. The activity of SpRY was validated in double stranded cleavage...assessment of the PAM compatibility and editing activities of Cas9 variants. Nucleic Acids Res. 2021 Sep...& Walton et al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver...several bps away from a desired site does not have the same devastating impact on efficiency as 10-15 bps away...
Showing: 141 - 160 of 356 results