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Overview
Product name Acetyl-Histone H3-K27 Rabbit mAb Catalog No. A22264 Host species Rabbit Purification method Affinity purification Isotype IgG CloneNo. ARC54943 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 acetylated 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 Calculated MW 15kDa Observed MW 17kDa Applications
Reactivity Human, Mouse, Rat, Other (Wide Range) Tested applications WB IHC-P IF/ICC IP ChIP ChIP-seq DB Recommended dilution - DB 1:2000 - 1:20000
- WB 1:2000 - 1:20000
- IHC-P 1:50 - 1:200
- IF/ICC 1:50 - 1:200
- ChIP 1:50 - 1:200
- ChIP-seq 1:50 - 1:200
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 TSA, NIH/3T3 TSA, C6 TSA Cellular location Chromosome, Nucleus 
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3-K27 Rabbit mAb (A22264). The ChIP sequencing results indicate the enrichment pattern of Acetyl-Histone H3-K27 in selected genomic region and representative gene loci (RPL30), as shown in figure.

Western blot analysis of HeLa, using Acetyl-Histone H3-K27 antibody (A22264) at 1:20000 dilution.HeLa cells were treated by TSA (1 uM) at 37℃ for 18 hours.
Secondary antibody: HRP 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: 30s.
Western blot analysis of NIH/3T3, using Acetyl-Histone H3-K27 antibody (A22264) at 1:20000 dilution.NIH/3T3 cells were treated by TSA (1 uM) at 37℃ for 18 hours.
Secondary antibody: HRP 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: 30s.
Western blot analysis of C6, using Acetyl-Histone H3-K27 antibody (A22264) at 1:20000 dilution.C6 cells were treated by TSA (1 uM) at 37℃ for 18 hours.
Secondary antibody: HRP 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: 30s.
Dot-blot analysis of all sorts of peptides using Acetyl-Histone H3-K27 antibody (A22264) at 1:2000 dilution.

Immunohistochemistry analysis of paraffin-embedded human lung using Acetyl-Histone H3-K27 Rabbit mAb (A22264) at dilution of 1:100 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded human spleen using Acetyl-Histone H3-K27 Rabbit mAb (A22264) at dilution of 1:100 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded rat intestine using Acetyl-Histone H3-K27 Rabbit mAb (A22264) at dilution of 1:100 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunofluorescence analysis of HeLa treated by TSA and HeLa cells using Acetyl-Histone H3-K27 Rabbit mAb (A22264) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using Acetyl-Histone H3-K27 antibody (A22264) 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.
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Key applications Western blotting, Immunohistochemistry, Immunofluorescence Host species Кролик Antibody type Primary Antibodies Reactivity Человек, Мышь, Крыса, Другое (Широкий ассортимент) Immunogen A synthetic acetylated peptide around K27 of human Histone H3 (NP_003520.1). Isotype IgG







