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Acetyl-Histone H3-K56 Rabbit mAb (A22565)

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)

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Acetyl-Histone H3-K56 mAb (A22565). The ChIP sequencing results indicate the enrichment pattern of Acetyl-Histone H3-K56 in selected genomic region and representative gene loci (GAPDH), as shown in figure.

ABclonal: - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina(RK20265) from 10⁵ K562 cells with 1μg Acetyl-Histone H3-K56 (A22565) , along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of Acetyl-Histone H3-K56 in representative gene loci (GAPDH), as shown in figure.

ABclonal:Western blot - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

Western blot analysis of lysates from NIH/3T3 cells, using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at 1:7000 dilution. NIH/3T3 cells were treated by TSA (1 uM) at 37℃ for 18 hours.
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: 0.5s.

Western blot analysis of lysates from C6 cells, using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at 1:7000 dilution. C6 cells were treated by TSA (1 uM) at 37℃ for 18 hours.
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: 0.5s.

Western blot analysis of lysates from Hela cells, using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at 1:7000 dilution. Hela cells were treated by TSA (1 uM) at 37℃ for 18 hours.
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:Dot Blot - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

Dot-blot analysis of all sorts of peptides using Acetyl-Histone H3-K56 antibody (A22565) at 1:10000 dilution.

ABclonal:Immunohistochemistry - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human esophagus tissue using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

Confocal imaging of HeLa cells (treated with TSA) and HeLa cells (untreated) using Acetyl-Histone H3-K56 Rabbit mAb (A22565, 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.

Immunofluorescence analysis of NIH-3T3 treated with TSA and NIH-3T3 cells using Acetyl-Histone H3-K56 Rabbit mAb (A22565) at dilution of 1:50 (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 - Acetyl-Histone H3-K56 Rabbit mAb (A22565)

Chromatin immunoprecipitation analysis of extracts of NIH/3T3 cells, using Acetyl-Histone H3-K56 antibody (A22565) 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.

Overview

Product name	Acetyl-Histone H3-K56 Rabbit mAb
Catalog No.	A22565
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC55111

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 K56 of human Histone H3 (NP_003520.1).
Sequence	MARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEACEAY
Gene ID
Swiss Prot
Synonyms	H3.4; H3/g; H3FT; H3t; HIST3H3; Histone H3; HIST1H3A; Acetyl-Histone H3-K56
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 1:2000 - 1:8000 DB 1:2000 - 1:10000 IHC-P 1:1000 - 1:5000 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 5μg antibody for 5μg-10μg of Chromatin ChIP-seq 1:50 - 1:200 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 treated by TSA, NIH/3T3 treated by TSA, C6 treated by TSA
Cellular location	Chromosome, nucleus.
Customer validation	WB(Mus musculus)