Name: Anti-EHMT2/G9A antibody [EPR18894]
Brand: RabMAb
SKU: ab185050
Rating: 5 (4 reviews)

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製品名

Anti-EHMT2/G9A antibody [EPR18894]
EHMT2/G9A 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR18894] to EHMT2/G9A
由来種

Rabbit
アプリケーション

適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
種交差性

交差種: Mouse, Rat, Human
免疫原

Recombinant fragment within Human EHMT2/G9A aa 1-300. The exact sequence is proprietary.
Database link: Q96KQ7

Run BLAST with Run BLAST with
ポジティブ･コントロール
- WB: HEK-293, Jurkat, HepG2, NCCIT, F9, Neuro-2a, LLC1, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. Human fetal heart and fetal kidney lysates. IHC-P: Human colon, Human gastric adenocarcinoma, mouse liver and rat kidney tissues. ICC/IF: HeLa. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
特記事項

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
バッファー

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR18894
アイソタイプ

IgG
研究分野

Our Abpromise guarantee covers the use of ab185050 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション	Abreviews	特記事項
WB		1/1000. Detects a band of approximately 170, 160 kDa (predicted molecular weight: 132 kDa).
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/1000.
Flow Cyt		1/150.
IP		1/50.

機能

Histone methyltransferase that specifically mono- and dimethylates 'Lys-9' of histone H3 (H3K9me1 and H3K9me2, respectively) in euchromatin. H3K9me represents a specific tag for epigenetic transcriptional repression by recruiting HP1 proteins to methylated histones. Also mediates monomethylation of 'Lys-56' of histone H3 (H3K56me1) in G1 phase, leading to promote interaction between histone H3 and PCNA and regulating DNA replication. Also weakly methylates 'Lys-27' of histone H3 (H3K27me). Also required for DNA methylation, the histone methyltransferase activity is not required for DNA methylation, suggesting that these 2 activities function independently. Probably targeted to histone H3 by different DNA-binding proteins like E2F6, MGA, MAX and/or DP1. May also methylate histone H1. In addition to the histone methyltransferase activity, also methylates non-histone proteins: mediates dimethylation of 'Lys-373' of p53/TP53. Also methylates CDYL, WIZ, ACIN1, DNMT1, HDAC1, ERCC6, KLF12 and itself.
組織特異性

Expressed in all tissues examined, with high levels in fetal liver, thymus, lymph node, spleen and peripheral blood leukocytes and lower level in bone marrow.
配列類似性

Belongs to the class V-like SAM-binding methyltransferase superfamily. Histone-lysine methyltransferase family. Suvar3-9 subfamily.
Contains 7 ANK repeats.
Contains 1 post-SET domain.
Contains 1 pre-SET domain.
Contains 1 SET domain.
ドメイン

The SET domain mediates interaction with WIZ.
The ANK repeats bind H3K9me1 and H3K9me2.
翻訳後修飾

Methylated at Lys-185; automethylated.
細胞内局在

Nucleus. Chromosome. Associates with euchromatic regions. Does not associate with heterochromatin.
Target information above from: UniProt accession Q96KQ7 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 10919 Human
- Entrez Gene: 110147 Mouse
- Entrez Gene: 361798 Rat
- Omim: 604599 Human
- SwissProt: Q96KQ7 Human
- SwissProt: Q9Z148 Mouse
- Unigene: 709218 Human
- Unigene: 35345 Mouse
別名
- Ankyrin repeat containing protein antibody
- Bat 8 antibody
- Bat8 antibody
- C6orf30 antibody
- DKFZp686H08213 antibody
- EHMT 2 antibody
- Ehmt2 antibody
- EHMT2_HUMAN antibody
- Euchromatic histone lysine methyltransferase 2 antibody
- Euchromatic histone lysine N methyltransferase 2 antibody
- Euchromatic histone-lysine N-methyltransferase 2 antibody
- FLJ35547 antibody
- G 9a antibody
- G9 a antibody
- G9a protein antibody
- G9A antibody
- G9A histone methyltransferase antibody
- GAT8 antibody
- H3 K9 HMTase 3 antibody
- H3-K9-HMTase 3 antibody
- Histone H3 K9 methyltransferase 3 antibody
- Histone H3 K9 methyltransferase3 antibody
- Histone H3-K9 methyltransferase 3 antibody
- Histone lysine N methyltransferase antibody
- Histone lysine N methyltransferase EHMT2 antibody
- Histone lysine N methyltransferase, H3 lysine 9 specific 3 antibody
- Histone lysine N methyltransferase, H3 lysine 9 specific3 antibody
- Histone-lysine N-methyltransferase EHMT2 antibody
- HLA B associated transcript 8 antibody
- HLA-B-associated transcript 8 antibody
- KMT 1C antibody
- KMT1 C antibody
- Lysine N methyltransferase 1C antibody
- Lysine N-methyltransferase 1C antibody
- NG 36 antibody
- NG36 antibody
- Protein G9a antibody
see all

Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: EHMT2/G9A knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab185050 observed at 160 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab185050 was shown to recognize EHMT2/G9A when EHMT2/G9A knockout samples were used, along with additional cross-reactive bands. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on rat kidney is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling EHMT2/G9A with ab185050 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab185050 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 4 : NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 132 kDa
Observed band size: 160,170 kDa
why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution + Human fetal heart lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 132 kDa
Observed band size: 160,170 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 132 kDa
Observed band size: 160,170 kDa why is the actual band size different from the predicted?

Exposure time: 8 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

Lane 1 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 2 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate
Lane 3 : LLC1 (Mouse lung carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 132 kDa
Observed band size: 160,170 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
Western blot - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

All lanes : Anti-EHMT2/G9A antibody [EPR18894] (ab185050) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 132 kDa
Observed band size: 160,170 kDa why is the actual band size different from the predicted?

Exposure time: 8 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on epithelial cells of the normal Human colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on tumor cells of the gastric adenocarcinoma is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling EHMT2/G9A with ab185050 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nucleus staining on hepatocytes of the mouse liver is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling EHMT2/G9A with ab185050 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing no staining on MCF7 cell line, as MCF7 cells have a very low level expression of EHMT2/G9A.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab185050 at 1/70 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Flow Cytometry - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling EHMT2/G9A with ab185050 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-EHMT2/G9A antibody [EPR18894] (ab185050)

EHMT2/G9A was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185050 at 1/50 dilution.
Western blot was performed from the immunoprecipitate using ab185050 at 1/1000 dilution.
VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10ug (Input).
Lane 2: ab185050 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185050 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

This product has been referenced in:

Rowbotham SP et al. H3K9 methyltransferases and demethylases control lung tumor-propagating cells and lung cancer progression. Nat Commun 9:4559 (2018). Read more (PubMed: 30455465) »

See 1 Publication for this product

ab185050 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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レビューを送る質問を送る

1-4 of 4 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Human Dermal Microvascular Endothelial Cells)

Permeabilization

Yes - 0.1% Triton-X100

Specification

Human Dermal Microvascular Endothelial Cells

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

投稿 Jul 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunoprecipitation

Sample

Human Cell lysate - whole cell (Dermal microvascular endothelium)

Total protein in input

200 µg

Immuno-precipitation step

Protein A/G

Specification

Dermal microvascular endothelium

Abcam user community

Verified customer

投稿 Apr 20 2017

Abcam has not validated the combination of species/application used in this Abreview.

Application

ChIP

Sample

Human Cell lysate - nuclear (B Cells)

Negative control

Isotype IgG

Specification

B Cells

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Methanol free Formaldehyde

Positive control

Gene promoter specific primers

Abcam user community

Verified customer

投稿 Mar 17 2017

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - nuclear (Human Dermal Microvascular Endothelial Cells)

Gel Running Conditions

Reduced Denaturing (7.5%)

Loading amount

20 µg

Specification

Human Dermal Microvascular Endothelial Cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Feb 28 2017

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

カスタム・ソリューション & パートナーシップ

Anti-EHMT2/G9A 抗体 [EPR18894] (ab185050)

製品の概要

製品の特性

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アプリケーション

ターゲット情報

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データシートおよび資料

参考文献

This product has been referenced in:

レビューと Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-EHMT2/G9A antibody [EPR18894]

Immunoprecipitation abreview for Anti-EHMT2/G9A antibody [EPR18894]

ChIP abreview for Anti-EHMT2/G9A antibody [EPR18894]

Western blot abreview for Anti-EHMT2/G9A antibody [EPR18894]