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Histone H3 (di methyl K27)
Histone H3 (di methyl K27) Recombinant Rabbit Monoclonal Antibody [PSH05-88]

Histone H3 (di methyl K27) Recombinant Rabbit Monoclonal Antibody [PSH05-88]

DATA SHEET
MSDS
COA

Usd: 385

Specification

100μl

Quantity

Bulk Order Quote | Customization Request

Catalog# HA722486

Histone H3 (di methyl K27) Recombinant Rabbit Monoclonal Antibody [PSH05-88]

Application

WB
IF-Cell
ChIP

Reactivity

Human
Mouse
Rat

BSA and Azide free

HA751035
不含抗保成分

Conjugation

unconjugated

Overview

Product Name

Histone H3 (di methyl K27) Recombinant Rabbit Monoclonal Antibody [PSH05-88]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is di-methylated.

Product Specificity

Di-Methyl-Histone H3 (Lys27) (PSH05-88) Rabbit mAb detects endogenous levels of histone H3 when di-methylated on Lys27. The antibody does not cross-react with mono-methylated or tri-methylated Lys27.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, ChIP

Molecular Weight

Predicted band size: 15 kDa

Positive Control

HeLa cell lysate, SH-SY5Y cell lysate, HCT 116 cell lysate, C6 cell lysate, MCF7 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, MCF7, NIH/3T3, C6.

Conjugation

unconjugated

Clone Number

PSH05-88

Product Features

Form

Liquid

Concentration

Lot Concentration Lookup

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:1,000

IF-Cell
1:100

ChIP
Use 0.5~2 μg for 25 μg of chromatin.

Target

Function

The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination. Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development. Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing. Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5). The discovery of histone demethylases, such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1, has shown that methylation is a reversible epigenetic marker.

Background References

1. Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.

2. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop, 1-27.

3. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.

4. Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.

5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.

6. Shi, X. et al. (2006) Nature 442, 96-9.

7. Wysocka, J. et al. (2006) Nature 442, 86-90.

8. Wysocka, J. et al. (2005) Cell 121, 859-72.

9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P68431 Human

SwissProt: P84243 Human

SwissProt: Q16695 Human

SwissProt: Q6NXT2 Human

SwissProt: Q71DI3 Human

SwissProt: P68433 Mouse

SwissProt: P84228 Mouse

SwissProt: Q6LED0 Rat

Synonyms

H3 histone family member E pseudogene antibody

H3 histone family, member A antibody

H3/A antibody

H31_HUMAN antibody

H3F3 antibody

H3FA antibody

Hist1h3a antibody

HIST1H3B antibody

HIST1H3C antibody

HIST1H3D antibody

Expand

H3 histone family member E pseudogene antibody

H3 histone family, member A antibody

H3/A antibody

H31_HUMAN antibody

H3F3 antibody

H3FA antibody

Hist1h3a antibody

HIST1H3B antibody

HIST1H3C antibody

HIST1H3D antibody

HIST1H3E antibody

HIST1H3F antibody

HIST1H3G antibody

HIST1H3H antibody

HIST1H3I antibody

HIST1H3J antibody

HIST3H3 antibody

histone 1, H3a antibody

Histone cluster 1, H3a antibody

Histone H3 3 pseudogene antibody

Histone H3.1 antibody

Histone H3/a antibody

Histone H3/b antibody

Histone H3/c antibody

Histone H3/d antibody

Histone H3/f antibody

Histone H3/h antibody

Histone H3/i antibody

Histone H3/j antibody

Histone H3/k antibody

Histone H3/l antibody

H3K27me2

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Images

☑ Cell treatment (CT)

Western blot analysis of Histone H3 (di methyl K27) on different lysates with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: SH-SY5Y cell lysate
Lane 3: HCT 116 cell lysate
Lane 4: C6 cell lysate
Lane 5: MCF7 cell lysate
Lane 6: MCF7 treated with 10μM EED226 for 72 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722486) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Histone H3 (di methyl K27) on different lysates with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/1,000 dilution.

Lane 1: RAW264.7 cell lysate (20 µg/Lane)
Lane 2: C2C12 cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (20 µg/Lane)

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 12 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722486) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells with Histone H3 (di methyl K27) (HA722486) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Immunocytochemistry analysis of MCF7 cells labeling Histone H3 (di methyl K27) with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H3 (di methyl K27) with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of C6 cells labeling Histone H3 (di methyl K27) with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (di methyl K27) antibody (HA722486) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.