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KMT6 / EZH2 Recombinant Rabbit Monoclonal Antibody [PSH04-14] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750937

KMT6 / EZH2 Recombinant Rabbit Monoclonal Antibody [PSH04-14] - BSA and Azide free

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

KMT6 / EZH2 Recombinant Rabbit Monoclonal Antibody [PSH04-14] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human EZH2 aa 201-250 / 746.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 85 kDa

Positive Control

HEK-293 cell lysate, 293T cell lysate, MCF7 cell lysate, HeLa cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, C6 cell lysate, HeLa, Neuro-2a.

Conjugation

unconjugated

Clone Number

PSH04-14

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

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. Displays a preference for substrates with less methylation, loses activity when progressively more methyl groups are incorporated into H3K27, H3K27me0 > H3K27me1 > H3K27me2. Compared to EZH1-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 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes. Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-ARNTL/BMAL1 heterodimer; involved in the di and trimethylation of 'Lys-27' of histone H3 on PER1/2 promoters which is necessary for the CRY1/2 proteins to inhibit transcription.

Background References

1. Fu TG et al. miR-143 inhibits oncogenic traits by degrading NUAK2 in glioblastoma. Int J Mol Med 37:1627-35 (2016).

2. Choi HJ et al. Significance of EZH2 expression in canine mammary tumors. BMC Vet Res 12:164 (2016).

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q15910 Human

SwissProt: Q61188 Mouse

Entrez Gene: 312299 Rat

Synonyms

Enhancer of zeste 2 antibody

enhancer of zeste 2 polycomb repressive complex 2 subunit antibody

Enhancer of zeste homolog 2 (Drosophila) antibody

Enhancer of zeste homolog 2 antibody

Enhancer of zeste, Drosophila, homolog 2 antibody

ENX 1 antibody

Enx 1h antibody

ENX-1 antibody

ENX1 antibody

Enx1h antibody

Expand

Images

  • Western blot analysis of KMT6 / EZH2 on different lysates with Rabbit anti-KMT6 / EZH2 antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: 293T cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: HeLa cell lysate<br />Lane 5: COS-1 cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: PC-12 cell lysate<br />Lane 8: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of KMT6 / EZH2 on different lysates with Rabbit anti-KMT6 / EZH2 antibody (HA750937) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: 293T cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: HeLa cell lysate
    Lane 5: COS-1 cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: PC-12 cell lysate
    Lane 8: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 85 kDa
    Observed band size: 85 kDa

    Exposure time: 3 minutes;

    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 (HA750937) 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.

  • Immunocytochemistry analysis of HeLa cells labeling KMT6 / EZH2 with Rabbit anti-KMT6 / EZH2 antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/100 dilution.<br /><br />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-KMT6 / EZH2 antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling KMT6 / EZH2 with Rabbit anti-KMT6 / EZH2 antibody (HA750937) 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-KMT6 / EZH2 antibody (HA750937) 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 Neuro-2a cells labeling KMT6 / EZH2 with Rabbit anti-KMT6 / EZH2 antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/100 dilution.<br /><br />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-KMT6 / EZH2 antibody (<a href="/products/HA750937" style="font-weight: bold;text-decoration: underline;">HA750937</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Neuro-2a cells labeling KMT6 / EZH2 with Rabbit anti-KMT6 / EZH2 antibody (HA750937) 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-KMT6 / EZH2 antibody (HA750937) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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