EHMT2/G9A Recombinant Rabbit mAb (S-4200)

Catalog Number: S0B6612 Application: WB, IHC-P, ICC, IF, ChIP Reactivity: Hu, Ms, Rt, Mk Conjugation: Unconjugated Brand: Starter

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$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	EHMT2/G9A
Synonyms	Histone-lysine N-methyltransferase EHMT2; Euchromatic histone-lysine N-methyltransferase 2; HLA-B-associated transcript 8; Histone H3-K9 methyltransferase 3 (H3-K9-HMTase 3); Lysine N-methyltransferase 1C; Protein G9a; BAT8; C6orf30; KMT1C; NG36
Location	Nucleus
Accession	Q96KQ7
Clone Number	S-4200
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, IHC-P, ICC, IF, ChIP
Reactivity	Hu, Ms, Rt, Mk
Positive Sample	MCF7, HL-60, HEK-293, HT-1080, COS-7
Purification	Protein A
Concentration	0.5 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:1000-1:2000	Hu, Mk
IHC-P	1:1000	Hu, Ms, Rt
ICC	1:500	Hu
IF	1:500	Hu
ChIP	1:20-1:50	Hu

Background

EHMT2 (also called G9a) is a nuclear lysine methyltransferase that chiefly di-methylates histone H3 at lysine 9 (H3K9me2) and lysine 27 (H3K27me1/2) to establish facultative heterochromatin and transcriptionally silence target genes, and it can act either as a repressor or, through auto-methylation-dependent recruitment of cofactors such as HP1γ and p300, as a co-activator of transcription; the enzyme exists as a homodimer or heterodimer with the related GLP/EHMT1, contains an N-terminal ankyrin-repeat domain for chromatin docking and a catalytic SET domain for methyl transfer, and its dysregulation—via overexpression in diverse solid and hematologic cancers or down-regulation in addiction and neuropathic pain—alters the expression of hundreds of genes by both histone and non-histone methylation, thereby controlling embryonic development, stem-cell lineage commitment, immune-cell differentiation, synaptic remodeling, and tumorigenicity .

Picture

Western Blot

WB result of EHMT2/G9A Recombinant Rabbit mAb
Primary antibody: EHMT2/G9A Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: MCF7 whole cell lysate 20 µg
Lane 2: HL-60 whole cell lysate 20 µg
Lane 3: HEK-293 whole cell lysate 20 µg
Lane 4: HT-1080 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 132 kDa
Observed MW: 130-180 kDa

WB result of EHMT2/G9A Recombinant Rabbit mAb
Primary antibody: EHMT2/G9A Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 132 kDa
Observed MW: 180 kDa

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human colon. Anti-EHMT2/G9A antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon cancer. Anti-EHMT2/G9A antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded mouse liver. Anti-EHMT2/G9A antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded rat kidney. Anti-EHMT2/G9A antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in HeLa cells. Anti-EHMT2/G9A antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).

Immunofluorescence

IF shows positive staining in paraffin-embedded human ovarian cancer. Anti- EHMT2/G9A antibody was used at 1/500 dilution (magenta) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 647) (S0B4005) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.

IF shows positive staining in paraffin-embedded human cervical cancer. Anti- EHMT2/G9A antibody was used at 1/500 dilution (magenta) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 647) (S0B4005) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.

ChIP

Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used EHMT2/G9A Recombinant Rabbit mAb (S-4200) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post-immunoprecipitation, both samples were washed, eluted, and cross-links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of VRK3, ZNF19 and SAT-α in
EHMT2/G9A Recombinant Rabbit mAb (S-4200) -immunoprecipitated
sample.