제품 > 항체 > 메틸화 특이적 항체

TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

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 K-562 cells with 1 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146), followed by incubation with Goat Anti-Rabbit IgG(H+L)(AS070). The CUT&Tag results denote the enrichment pattern of TriMethyl-Histone H3-K4 around RPL30 gene.

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina (RK20265) from 10⁵ K-562 cells with 1 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146), followed by incubation with Goat Anti-Rabbit IgG (H+L)(AS070). The CUT&Tag results denote the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).

ABclonal:ChIP-seq - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from 293T using 5 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K4 in the representative genomic region surrounding RPL30 gene.

Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from 293T using 5 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).

ABclonal:Western blot - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb (A22146)at 1:5000 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: 45s.

ABclonal:Immunofluorescence - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Confocal imaging of PC-12 cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of NIH/3T3 cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of HeLa cells using TriMethyl-Histone H3-K4 Rabbit mAb (A22146, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Dot Blot - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Dot-blot analysis of all sorts of peptides using TriMethyl-Histone H3-K4 antibody (A22146) at 1:50000 dilution.

ABclonal:Immunohistochemistry - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Immunohistochemistry analysis of paraffin-embedded Human liver cancer using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at dilution of 1:200 (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-K4 Rabbit mAb (A22146) at dilution of 1:200 (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 brain using TriMethyl-Histone H3-K4 Rabbit mAb (A22146) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Chromatin Immunoprecipitation - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K4 antibody (A22146) 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-K4 Rabbit mAb
Catalog No.	A22146
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC55095

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 K4 of human Histone H3 (NP_003520.1).
Sequence	MARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAVMALQEACEAY
Gene ID
Swiss Prot
Synonyms	H3.4; H3/g; H3FT; H3t; HIST3H3; Histone H3; HIST1H3A; TriMethyl-Histone H3-K4
Calculated MW	15kDa
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- 1:10000 DB 1:10000 - 1:60000 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 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, C6
Cellular location	Chromosome, nucleus.
Customer validation	(Arabidopsis Seedlings, Tilapia mossambica) WB(Homo sapiens, Other, Mus musculus) CRISPR(Homo sapiens) ChIP(Macaca mulatta)

Documents

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

Lot number

Research Area

TriMethyl-Histone H3-K4 Rabbit mAb images

- TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina (RK20265) from 10⁵ K-562 cells with 1 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146), followed by incubation with Goat Anti-Rabbit IgG (H+L)(AS070). The CUT&Tag results denote the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).

ChIP-seq - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from 293T using 5 μg of TriMethyl-Histone H3-K4 Rabbit mAb (A22146). DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2 (RK20228). The ChIP sequencing results indicate the enrichment pattern of TriMethyl-Histone H3-K4 across chromosome 8 (upper panel) and the genomic region encompassing RPL30, a representative gene enriched in TriMethyl-Histone H3-K4 (lower panel).

Western blot - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Immunofluorescence - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Dot Blot - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Dot-blot analysis of all sorts of peptides using TriMethyl-Histone H3-K4 antibody (A22146) at 1:50000 dilution.

Immunohistochemistry - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

Chromatin Immunoprecipitation - TriMethyl-Histone H3-K4 Rabbit mAb (A22146)

* For research use only. Not for therapeutic or diagnostic purposes.