Anti-MBD2 抗体 [EPR18361] (ab188474)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18361] to MBD2
- Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-MBD2 antibody [EPR18361]
MBD2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR18361] to MBD2 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IHC-P, ICC/IF, IP, WBmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: HeLa, NIH/3T3, MCF7, A-375 and PC-12 cell lysates; mouse brain, mouse heart and rat brain lysates. IHC-P: Human colon, human gastric cancer, mouse stomach and rat colon tissues. ICC/IF: HepG2 cells. IP: HeLa whole cell lysate.
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特記事項
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.
製品の特性
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR18361 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab188474の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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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 |
1/500.
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IP |
1/50.
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WB |
1/1000. Detects a band of approximately 43, 29 kDa (predicted molecular weight: 43 kDa).
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IP
1/50. |
WB
1/1000. Detects a band of approximately 43, 29 kDa (predicted molecular weight: 43 kDa). |
ターゲット情報
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機能
Binds CpG islands in promoters where the DNA is methylated at position 5 of cytosine within CpG dinucleotides. Binds hemimethylated DNA as well. Recruits histone deacetylases and DNA methyltransferases. Acts as transcriptional repressor and plays a role in gene silencing. Functions as a scaffold protein, targeting GATAD2A and GATAD2B to chromatin to promote repression. May enhance the activation of some unmethylated cAMP-responsive promoters. -
組織特異性
Highly expressed in brain, heart, kidney, stomach, testis and placenta. -
配列類似性
Contains 1 MBD (methyl-CpG-binding) domain. -
細胞内局在
Nucleus. Nuclear, in discrete foci. Detected at replication foci in late S phase. - Information by UniProt
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参照データベース
- Entrez Gene: 8932 Human
- Entrez Gene: 17191 Mouse
- Entrez Gene: 680172 Rat
- Omim: 603547 Human
- SwissProt: Q9UBB5 Human
- SwissProt: Q9Z2E1 Mouse
- Unigene: 25674 Human
- Unigene: 322 Mouse
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別名
- Demethylase antibody
- DMTase antibody
- MBD 2 antibody
see all
画像
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MBD2 knockout HAP1 cell lysate (20 µg)
Lane 3: A375 cell lysate (20 µg)
Lane 4: Human brain tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab188474 observed at 32 & 49 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab188474 was shown to specifically react with MBD2 when MBD2 knockout samples were used. Wild-type and MBD2 knockout samples were subjected to SDS-PAGE. ab188474 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 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 1/10000 dilution for 1 h at room temperature before imaging. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling MBD2 with ab188474 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab188474 at 1/250 dilution, followed by Goat Anti-Mouse (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. -
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat colon tissue 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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Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling MBD2 with purified ab188474 at 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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All lanes : Anti-MBD2 antibody [EPR18361] (ab188474) at 1/10000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 43 kDa
Observed band size: 29,43 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID: 17353267).
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human colon tissue tissue 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-MBD2 antibody [EPR18361] (ab188474) at 1/10000 dilution
Lane 1 : A-375 (Human malignant melanoma cell line) lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 43 kDa
Observed band size: 29,43 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes5% NFDM/TBST: Blocking and dilution buffer.
The observed MW is consistent with what has been described in the literature (PMID:17353267).
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Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human gastric cancer tissue 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-MBD2 antibody [EPR18361] (ab188474) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 43 kDa
Observed band size: 29,43 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:17353267).
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Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling MBD2 with ab188474 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse stomach tissue 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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MBD2 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab188474 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab188474 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate 10µg (Input).
Lane 2: ab188474 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188474 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (3)
ab188474 は 3 報の論文で使用されています。
- Yue T et al. MBD2 acts as a repressor to maintain the homeostasis of the Th1 program in type 1 diabetes by regulating the STAT1-IFN-? axis. Cell Death Differ N/A:N/A (2021). PubMed: 34420035
- Ge Y et al. Mbd2 Mediates Retinal Cell Apoptosis by Targeting the lncRNA Mbd2-AL1/miR-188-3p/Traf3 Axis in Ischemia/Reperfusion Injury. Mol Ther Nucleic Acids 19:1250-1265 (2020). PubMed: 32074940
- Gong W et al. Expression and clinical significance of methyl-CpG binding domain protein 2 in high-grade serous ovarian cancer. Oncol Lett 20:2749-2756 (2020). PubMed: 32782591