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TriMethyl-Histone H3-K27 Rabbit mAb (A22396)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat, Other (Wide Range Predicted)

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina(RK20265) from 10⁵ K562 cells with 1μg TriMethyl-Histone H3-K27 Rabbit mAb(A22396), along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of H3K27Me3 in representative gene loci (MYT1), as shown in figure.

ABclonal:Western blot - TriMethyl-Histone H3-K27 Rabbit mAb (A22396)

Western blot analysis of various lysates, using TriMethyl-Histone H3-K27 Rabbit mAb (A22396) at 1:140000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 90s.

ABclonal:Immunohistochemistry - TriMethyl-Histone H3-K27 Rabbit mAb (A22396)

Immunohistochemistry analysis of paraffin-embedded Human liver using TriMethyl-Histone H3-K27 Rabbit mAb (A22396) at dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human spleen using TriMethyl-Histone H3-K27 Rabbit mAb (A22396) at dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - TriMethyl-Histone H3-K27 Rabbit mAb (A22396)

Immunofluorescence analysis of HeLa cells using TriMethyl-Histone H3-K27 Rabbit mAb (A22396) at dilution of 1:600 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

ABclonal:Chromatin Immunoprecipitation - TriMethyl-Histone H3-K27 Rabbit mAb (A22396)

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K27 antibody (A22396) 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.

Review

Overview

Product name	TriMethyl-Histone H3-K27 Rabbit mAb
Catalog No.	A22396
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC54167

Background

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 information

Immunogen	A synthetic trimethylated peptide around K27 of human Histone H3 (NP_003520.1).
Sequence	MARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEACEAY
Gene ID
Swiss Prot
Synonyms	H3t; H3.4; H3/g; H3FT; H3C16; HIST3H3; TriMethyl-Histone H3-K27
Calculated MW	16kDa
Observed MW	17kDa

Applications

Reactivity	Human, Mouse, Rat, Other (Wide Range Predicted)
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:10000 - 1:160000 IHC-P 1:500 - 1:1000 IF/ICC 1:500 - 1:1000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements. ChIP 5μg antibody for 5μg-10μg of Chromatin CUT&Tag 10⁵ cells /1 μg
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	HeLa, HCT 116, NIH/3T3, C6
Cellular location	Chromosome, Nucleus.
Customer validation	(Tilapia mossambica) CUT&Tag(Mus musculus) WB(Mus musculus)