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DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

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 DiMethyl-Histone H3-K4 Rabbit mAb (A22143). The ChIP sequencing results indicate the enrichment pattern of DiMethyl-Histone H3-K4 in selected genomic region and representative gene loci (GAPDH), as shown in figure.

ABclonal: - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina(RK20265) from 10⁵ K562 cells with 1 μg DiMethyl-Histone H3-K4 Rabbit mAb (A22295), along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of DiMethyl-Histone H3-K4 in representative gene loci (GAPDH), as shown in figure.

ABclonal:Western blot - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Western blot analysis of various lysates, using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at1:3000 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: 10s.

ABclonal:Dot Blot - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Dot-blot analysis of all sorts of peptides using DiMethyl-Histone H3-K4 antibody (A22143) at 1:5000 dilution.

ABclonal:Immunohistochemistry - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Immunohistochemistry analysis of paraffin-embedded Human colon tissue using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at a dilution of 1:3000 (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 kidney tissue using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at a dilution of 1:3000 (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 liver cancer tissue using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at a dilution of 1:3000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at a dilution of 1:3000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using DiMethyl-Histone H3-K4 Rabbit mAb (A22143) at a dilution of 1:3000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Confocal imaging of HeLa cells using DiMethyl-Histone H3-K4 Rabbit mAb (A22143, dilution 1:200)(Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) (Green). DAPI was used for nuclear staining (blue). Objective: 100x.

ABclonal:Immunoprecipitation - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Immunoprecipitation analysis of 600 μg extracts of 293F cells using 5 μg DiMethyl-Histone H3-K4 antibody (A22143). Western blot was performed from the immunoprecipitate using DiMethyl-Histone H3-K4 antibody (A22143) at a dilution of 1:2000.

ABclonal:Chromatin Immunoprecipitation - DiMethyl-Histone H3-K4 Rabbit mAb (A22143)

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using DiMethyl-Histone H3-K4 antibody (A22143) 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	DiMethyl-Histone H3-K4 Rabbit mAb
Catalog No.	A22143
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC55489

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 dimethylated peptide around K4 of human Histone H3 (NP_003520.1).
Sequence	MARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEACEAY
Gene ID
Swiss Prot
Synonyms	H3t; H3.4; H3/g; H3FT; H3C16; HIST3H3; DiMethyl-Histone H3-K4
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 1:1000 - 1:5000 DB 1:2000 - 1:6000 IHC-P 1:1000 - 1:5000 IF/ICC 1:50 - 1:200 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells 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 Immunoprecipitation
Positive samples	HeLa, NIH/3T3, C6
Cellular location	Chromosome, nucleus.
Customer validation	WB(Other)