重组Anti-EHMT2/G9A抗体[EPR18894] - ChIP Grade (ab185050)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18894] to EHMT2/G9A - ChIP Grade
- Suitable for: ChIP-sequencing, WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-EHMT2/G9A抗体[EPR18894] - ChIP Grade
参阅全部 EHMT2/G9A 一抗 -
描述
兔单克隆抗体[EPR18894] to EHMT2/G9A - ChIP Grade -
宿主
Rabbit -
经测试应用
适用于: ChIP-sequencing, WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, 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 (intra): HeLa cells. IP: HeLa whole cell lysate. ChIP-seq: HeLa cells.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18894 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab185050于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ChIP-sequencing |
Use 8µg for 107 cells.
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WB | (1) |
1/1000. Detects a band of approximately 170, 160 kDa (predicted molecular weight: 132 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/1000.
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IP | (1) |
1/50.
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Flow Cyt (Intra) |
1/150.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
说明 |
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ChIP-sequencing
Use 8µg for 107 cells. |
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. |
IP
1/50. |
Flow Cyt (Intra)
1/150. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
靶标
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功能
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. -
组织特异性
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. -
序列相似性
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. -
结构域
The SET domain mediates interaction with WIZ.
The ANK repeats bind H3K9me1 and H3K9me2. -
翻译后修饰
Methylated at Lys-185; automethylated. -
细胞定位
Nucleus. Chromosome. Associates with euchromatic regions. Does not associate with heterochromatin. - Information by UniProt
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数据库链接
- 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
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别名
- Ankyrin repeat containing protein antibody
- Bat 8 antibody
- Bat8 antibody
see all
图片
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL. 2.5X10^5 of Human wild-type HeLa cell line (ab255928) or EHMT2 (G9A) knockout HeLa cell line (ab265149) were used along with 5µg of ab185050 [EPR18894]. Assay Quality Control was conducted using 5µg Anti-CTCF (ab188408) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EHMT2/G9A knockout HeLa cell lysate
Lane 3 : HEK-293 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 132 kDaLanes 1-4: Merged signal (red and green). Green - ab185050 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.
ab185050 Anti-EHMT2/G9A antibody [EPR18894] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
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Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 8 µg of ab185050 [EPR18894]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (ab185050) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : EHMT2/G9A knockout HAP1 cell lysate
Lane 3 : HEK293 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 132 kDaLanes 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.
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All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
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Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
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Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
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All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
-
All lanes : Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID 16702210).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (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.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling EHMT2/G9Awith 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.
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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 Detection Reagent (HRP) (ab131366) was used for detection 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.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (33)
ab185050 被引用在 33 文献中.
- Zhao L et al. BRD4-PRC2 represses transcription of T-helper 2-specific negative regulators during T-cell differentiation. EMBO J 42:e111473 (2023). PubMed: 36719036
- Mu YR et al. Role and mechanism of FOXG1-related epigenetic modifications in cisplatin-induced hair cell damage. Front Mol Neurosci 16:1064579 (2023). PubMed: 37181652
- Vini R et al. 27-Hydroxycholesterol represses G9a expression via oestrogen receptor alpha in breast cancer. J Cell Mol Med 27:2744-2755 (2023). PubMed: 37614064
- Fang Z et al. Depletion of G9A attenuates imiquimod-induced psoriatic dermatitis via targeting EDAR-NF-κB signaling in keratinocyte. Cell Death Dis 14:627 (2023). PubMed: 37739945
- Swaminathan J et al. Cross-Talk Between Histone Methyltransferases and Demethylases Regulate REST Transcription During Neurogenesis. Front Oncol 12:855167 (2022). PubMed: 35600406