Anti-Bmi1 (phospho T275) 抗体 [EPR19848] (ab213723)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19848] to Bmi1 (phospho T275)
- Suitable for: WB, Dot blot, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Bmi1 (phospho T275) antibody [EPR19848]
Bmi1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR19848] to Bmi1 (phospho T275) -
由来種
Rabbit -
アプリケーション
適用あり: WB, Dot blot, ICC/IF, IP, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, 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: HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (+/- exposure to UV light). Dot blot: Bmi1 (phospho T275) peptide. ICC/IF: HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (exposed to UV light) and U2OS cells. Flow Cyt (intra): HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1. IP: HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (exposed to UV light).
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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: 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
EPR19848 -
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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Isotype control
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Recombinant Protein
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Related Products
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ab213723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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WB |
1/2000. Predicted molecular weight: 37 kDa.
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Dot blot |
1/1000.
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ICC/IF |
1/100.
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IP |
1/30.
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Flow Cyt (Intra) |
1/500.
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特記事項 |
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WB
1/2000. Predicted molecular weight: 37 kDa. |
Dot blot
1/1000. |
ICC/IF
1/100. |
IP
1/30. |
Flow Cyt (Intra)
1/500. |
ターゲット情報
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機能
Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility. In the PRC1 complex, it is required to stimulate the E3 ubiquitin-protein ligase activity of RNF2/RING2. -
配列類似性
Contains 1 RING-type zinc finger. -
翻訳後修飾
Monoubiquitinated (By similarity). May be polyubiquitinated; which does not lead to proteasomal degradation. -
細胞内局在
Nucleus. Cytoplasm. - Information by UniProt
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参照データベース
- Entrez Gene: 100532731 Human
- Entrez Gene: 648 Human
- Entrez Gene: 12151 Mouse
- Omim: 164831 Human
- SwissProt: P35226 Human
- SwissProt: P25916 Mouse
- Unigene: 380403 Human
- Unigene: 289584 Mouse
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別名
- B lymphoma Mo MLV insertion region (mouse) antibody
- B lymphoma Mo MLV insertion region 1 homolog antibody
- Bmi 1 antibody
see all
画像
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All lanes : Anti-Bmi1 (phospho T275) antibody [EPR19848] (ab213723) at 1/2000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector, whole cell lysate
Lane 2 : HEK-293T transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector, whole cell lysate
Lane 3 : HEK-293T transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector, followed by treatment with 800 J/m² UV, then cultured in DMEM containing 10% FBS for 30 minutes, whole cell lysate
Lane 4 : HEK-293T transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector, followed by treatment with 800 J/m² UV, then cultured in DMEM containing 10% FBS for 30 minutes, 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
Developed using the ECL technique.
Predicted band size: 37 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and dilution buffer: 2% BSA/TBST
The phosphorylation of Bmi1 at T275 is increased by UV treatment.
The expression plasmids were kindly provided by Dr. Wei Guo from Tsinghua University.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector labeling Bmi1 (phospho T275) with ab213723 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector. No staining was observed in HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector.
The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (Alexa Fluor® 647) (ab195884) at 1/200 dilution (white).
The negative controls are as follows:
-ve control 1: PBS instead of primary antibody (HEK-293T transfected with wild-type BMi1), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
-ve control 2: PBS instead of primary antibody (HEK-293T transfected with Bmi1 T275A construct), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.Plasmids were kindly provided by Dr. Wei Guo from Tsinghua University.
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Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector (left), or DDDDK and mCherry-tagged mouse Bmi1 WT expression vector (right) labeling Bmi1 (phospho T275) with ab213723 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (black).
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077),at 1/2000 dilution was used as the secondary antibody.
The cells were gated on the mCherry positive population.
The plasmids were kindly provided by Dr. Wei Guo from Tsinghua University.
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Bmi1 (phospho T275) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector, followed by treatment with 800 J/m² UV, then cultured in DMEM containing 10% FBS for 30 minutes) whole cell lysate with ab213723 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213723 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HEK-293T (transfected / UV treated) whole cell lysate 10 μg (Input).
Lane 2: ab213723 IP in HEK-293T (transfected / UV treated) lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213723 in HEK-293T (transfected / UV treated) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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Dot blot analysis of Bmi1 (phospho T275) labeled with ab213723 at 1/1,000 dilution.
Lane 1: Bmi1 (phospho T275) peptide.
Lane 2: Bmi1 non-phospho peptide.
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell line) cells labeling Bmi1 (phospho T275) with ab213723 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased nuclear foci staining observed in UV-treated U-2 OS cells. The cells were treated with 50 J/m² UV, then cultured in McCoy's 5a media supplemented with 10% FBS for 2 hours.
The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
The negative controls are as follows:
-ve control 1: PBS instead of primary antibody (non-treated cells), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
-ve control 2: PBS instead of primary antibody (UV treated cells), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
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 (0)
ab213723 は論文での使用が確認できていません。