TriMethyl-Histone H3-K9 Monoclonal Antibody (CAB22295)

Product Name:	TriMethyl-Histone H3-K9 Monoclonal Antibody
Product Code:	CAB22295
Reactivity:	Human, Mouse, Rat, Other (Wide Range)
Applications:	Western blotting, Immunofluorescence
Host Species:	Rabbit
Purification Method:	Affinity purification
Isotype:	IgG
Clone No:	ARC54898

Reactivity:	Human, Mouse, Rat, Other (Wide Range)
Tested Applications:	WB, IF/ICC, IP, ChIP, ChIP-seq, ELISA, DB, CUT&Tag
Key Applications:	Western blotting, Immunofluorescence
Recommended Dilution:	WB 1:2000-1:20000, IF/ICC 1:50-1:200, ChIP 1:50-1:200, ChIP-seq 1:20-1:100, DB 1:2000-1:20000, CUT&Tag 10, ⁵ cells /1 μg
Storage Buffer:	Store at -20°C. Avoid freeze / thaw cycles.Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Positive Samples:	HeLa, C2C12 , C6
Cellular Location:	Chromosome, Nucleus

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.

Immunogen:	A synthetic trimethylated peptide around K9 of human Histone H3 (NP_003520.1).
Sequence:	MARTK QTARK STGGK APRKQ LATKA ARKSA PATGG VKKPH RYRPG TVALR EIRRY QKSTE LLIRK LPFQR LVREI AQDFK TDLRF QSSAV MALQE ACEAY
Synonyms:	H3.4, H3/g, H3FT, H3t, HIST3H3, Histone H3, HIST1H3A
Calculated MW:	15kDa
Observed MW:	17kDa

	Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and TriMethyl-Histone H3-K9 mAb (CAB22295). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K9 in selected genomic region and representative gene loci (ZNF404,GAPDH), as shown in figure.
	CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina from 10⁵ K562 cells with 1 μg TriMethyl-Histone H3-K9 Rabbit mAb (CAB22295), along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of TriMethyl-Histone H3-K9 in representative gene loci (MYT1), as shown in figure.
	Western blot analysis of various lysates, using TriMethyl-Histone H3-K9 Rabbit mAb (CAB22295) at 1:20000 dilution. Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (CABS014) at 1:10000 dilution. Lysates/proteins: 25μg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit (AbGn00020). Exposure time: 30s.
	Dot-blot analysis of all sorts of peptides using TriMethyl-Histone H3-K9 antibody (CAB22295) at 1:20000 dilution.
	Confocal imaging of HeLa cells using TriMethyl-Histone H3-K9 Rabbit mAb (CAB22295,dilution 1:100)(Red). The cells were counterstained with α-Tubulin Mouse mAb (CABC012,dilution 1:400) (Green). DAPI was used for nuclear staining (blue). Objective: 100x.
	Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K9 antibody (CAB22295) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.