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TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)

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)

Dot-blot analysis of H3K9me0, H3K9me1, H3K9me2, H3K9me3, H3K14me0, H3K14me1, H3K14me2, H3K14me3, H3K27me0, H3K27me1, H3K27me2, H3K27me3，H3K36me0, H3K36me1, H3K36me2, H3K36me3，H3K79me0, H3K79me1, H3K79me2, H3K79me3 using TriMethyl-Histone H3-K27 Rabbit PolymAb® .

ABclonal:Western blot - TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)

Western blot analysis of various lysates using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Negative control (NC): H3 prokaryotic protein
Exposure time: 5s.

ABclonal:Immunohistochemistry - TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)

Immunohistochemistry analysis of paraffin-embedded Rat brain tissue using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer(pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer(pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer(pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human liver cancer tissue using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer(pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human breast tissue using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer(pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)

Confocal imaging of HeLa cells using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of C6 cells using TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Chromatin Immunoprecipitation - TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM)

Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa, using 2 μg of TriMethyl-Histone H3-K27 Rabbit PolymAb® (A25095PM) and Rabbit Control IgG (AC005). The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.

Overview

Product name	TriMethyl-Histone H3-K27 Rabbit PolymAb®
Catalog No.	A25095PM
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

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 1:500 - 1:1000 DB 1:500 - 1:1000 IHC-P 1:50 - 1:200 IF/ICC 1:50 - 1:200 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements. ChIP 2 μg antibody for 5μg-10μg of Chromatin
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
Cellular location	Chromosome, Nucleus.