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[K9-trimethyl]Histone H3 monoclonal antibody (6F12-H4)

 
ALX-804-673-C050 50 µg 291.00 USD
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Product Specification

Alternative Name:H3K9me3, Trimethyl-histone H3 (Lys9)
 
Clone:6F12-H4
 
Host:Mouse
 
Isotype:IgG1
 
Immunogen:Branched peptide of the sequence (QTARK(Me)3STGGKA)2-KC.
 
UniProt ID:P68431 (human Histone H3.1), Q71DI3 (human Histone H3.2), P84243 (human Histone H3.2)
 
Species reactivity:Human, Mouse
 
Specificity:Recognizes K9-trimethylated histone H3.
 
Crossreactivity:Cross-reacts with K9-dimethylated and weakly with K9 -monomethylated histone H3. Does not cross-react with unmethylated histone H3.
 
Applications:ELISA, ICC, IP, WB
ChIP, Dot Blot
 
Formulation:Liquid. In PBS containing 0.02% sodium azide.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Blue Ice
 
Short Term Storage:+4°C
 
Long Term Storage:-20°C
 
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Figure 1: Western blot analysis of HeLa histones using MAb to Histone H3 (K9 methylated) (6F12-H4) (Prod. No. ALX-804-673).Lane 1: Fibercell supernatant (1:500)Lane 2: Purified antibody (ALX-804-673) (1:1’000)Lane 3: Purified antibody (ALX-804-673) (1:5’000)~1μg of HeLa cell total histones per lane, 10s exposure.
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Figure 2: Immunocytochemistry of irradiated mouse embryonic fibroblasts (iMEFs) stained with DAPI and MAb to Histone H3 (K9 methylated) (6F12-H4) (Prod. No. ALX-804-673).Figure courtesy of Susanne Opravil and Thomas Jenuwein, Institute of Molecular Pathology (IMP) Vienna, Austria
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Figure 3: ChIP analysis of known chromosomal Suv39h targets (H2K9me3 in major satellites, mouse ES cells) using MAb to Histone H3 (K9 methylated) (6F12-H4) (Prod. No. ALX-804-673).Figure courtesy of Inti Alberto De La Rosa Velazquez and Thomas Jenuwein, Institute of Molecular Pathology (IMP) Vienna, Austria.
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Figure 4: Dot-blot analysis using MAb to Histone H3 (K9 methylated) (6F12-H4) (Prod. No. ALX-804-673).Figure courtesy of Susanne Opravil and Thomas Jenuwein, Institute of Molecular Pathology (IMP) Vienna, Austria.
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Figure 5: Peptide competition assay using MAb to Histone H3 (K9 methylated) (6F12-H4) (Prod. No. ALX-804-673).Method: To determine the specificity of MAb to Histone H3 (K9 methylated) (6F12-H4), the antibody was diluted 1:200 in 0.5% NFDM in PBS-T and incubated for 6 hours at 4°C with a molar excess of peptides corresponding to aa 1-20 of human histone H3, containing either unmodified K9 (lanes 2-6), dimethylated K9 (lanes 8-12), or tri-methylated K9 (lanes 14-18). In each case, the molar excess of peptide over antibody was 100 fold, 50 fold, 20 fold, 10 fold, and 5 fold, respectively, as indicated by the arrowheads (from left to right) above the blot. The membrane containing histones prepared from HeLa cells was then incubated o/n at 4°C with the antibody/peptide solutions, and processed for Western blotting. As control, one aliquot of antibody was incubated with water only (lane 1). Lanes 7 and 13 were left empty.
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