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Render Timestamp: 2024-12-05T11:58:05.794Z
Commit: 224419269841c11382c4555dbee545259bf6c379
XML generation date: 2024-09-20 06:19:38.822
Product last modified at: 2024-11-26T17:45:09.495Z
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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751

Filter:
  • WB
  • IHC
  • IF
  • F
  • ChIP
  • C&R
  • C&T

    Supporting Data

    REACTIVITY H M R Mk Dm Sc
    SENSITIVITY Endogenous
    MW (kDa) 17
    Source/Isotype Rabbit IgG
    Application Key:
    • WB-Western Blotting 
    • IHC-Immunohistochemistry 
    • IF-Immunofluorescence 
    • F-Flow Cytometry 
    • ChIP-Chromatin Immunoprecipitation 
    • C&R-CUT & RUN 
    • C&T-CUT & Tag 
    Species Cross-Reactivity Key:
    • H-Human 
    • M-Mouse 
    • R-Rat 
    • Mk-Monkey 
    • Dm-D. melanogaster 
    • Sc-S. cerevisiae 

    Product Information

    Product Usage Information

    For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

    The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

    The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

    Application Dilution
    Western Blotting 1:1000
    Immunohistochemistry (Paraffin) 1:1000 - 1:4000
    Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
    Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
    Chromatin IP 1:50
    Chromatin IP-seq 1:50
    CUT&RUN 1:50
    CUT&Tag 1:50

    Storage

    Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

    For a carrier free (BSA and azide free) version of this product see product #19776.

    Protocol

    Specificity / Sensitivity

    Tri-Methyl-Histone H3 (Lys4) Antibody detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, the antibody does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20.

    Species Reactivity:

    Human, Mouse, Rat, Monkey, D. melanogaster, S. cerevisiae

    The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.

    Species predicted to react based on 100% sequence homology:

    Xenopus, Zebrafish

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys4 is tri-methylated.

    Background

    The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases, such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1, has shown that methylation is a reversible epigenetic marker (9).
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