EZH2 Monoclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (EZH2) Monoclonal Antibody, Unconjugated (bsm-51468M) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (EZH2) Monoclonal Antibody, Unconjugated (bsm-51468M) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.

$Jurkat cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with EZH2 Antibody(bsm-51468M)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).$

Jurkat cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with EZH2 Antibody(bsm-51468M)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).

$25 ug total protein per lane of various lysates (see on figure) probed with EZH2 monoclonal antibody, unconjugated (bsm-51468M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with EZH2 monoclonal antibody, unconjugated (bsm-51468M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Applications

WB
Flow-Cyt
ICC/IF

Reactivity

Human

Overview
Catalog #	bsm-51468m
Product Name	EZH2 Monoclonal Antibody
Applications	WB, Flow-Cyt, ICC/IF
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Mouse
Source	Recombinant human EZH2.
Clonality	Monoclonal
Clone #	E3H5
Concentration	0.5ug/ul
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	2146
Swiss Prot	Q15910
Subcellular location	Nucleus
Synonyms	Enhancer of zeste homolog 2; Enx1h; MGC9169; Enhancer of zeste 2; ENX-1; ENX 1; ENX1; EZH 2; EZH2_HUMAN; Histone-lysine N-methyltransferase EZH2; KMT6A; Lysine N-methyltransferase 6; Enhancer of zeste homolog 2 (Drosophila); Enhancer of zeste, Drosophila, homolog 2; KMT 6;KMT6A; KMT6; WVS2.
Background	Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4.
Application Dilution
WB	1:500-2000
Flow-Cyt	Flow-Cyt1ug/Test
ICC/IF	ICC/IF1:50-200