Anti-BNIP3 抗体 [EPR4034] (ab109362)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4034] to BNIP3
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-BNIP3 antibody [EPR4034]
BNIP3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR4034] to BNIP3 -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IFmore 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: Jurkat cells, Jurkat cells treated with etoposide, SH-SY5Y cells, SH-SY5Y cells treated with camptothecin and MCF-7 cells treated with Cocl2 lysates and rat kidney, mouse spleen and mouse kidney tissue lysates. IHC-P: Human renal adenocarcinoma and kidney tissues. ICC/IF: HeLa and SH-SY5Y cells.
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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. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 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
EPR4034 -
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
- Mouse kidney normal tissue lysate - total protein (ab29305)
- MCF7 whole cell lysate (ab29537)
- Jurkat whole cell lysate (ab30128)
- Mouse kidney tissue lysate - total protein (0 days) (ab7261)
- Mouse kidney tissue lysate (14 days) (ab7262)
- Mouse kidney tissue lysate (7 days) (ab7263)
- Jurkat whole cell lysate (ab7899)
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ab109362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 | (3) |
1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 22 kDa).
For unpurified use at 1/1000 - 1/10000. |
IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. (Heat to 98°C, allow to cool for 10-20 minutes). For unpurified use at 1/50 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/150.
For unpurified use at 1/100 - 1/250. |
特記事項 |
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WB
1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 22 kDa). For unpurified use at 1/1000 - 1/10000. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. (Heat to 98°C, allow to cool for 10-20 minutes). For unpurified use at 1/50 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/150. For unpurified use at 1/100 - 1/250. |
ターゲット情報
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機能
Apoptosis-inducing protein that, which can overcome BCL2 suppression. May play a role in repartitioning calcium between the two major intracellular calcium stores in association with BCL2. -
配列類似性
Belongs to the NIP3 family. -
細胞内局在
Mitochondrion. Mitochondrion membrane. Coexpression with the EIB 19-kDa protein results in a shift in NIP3 localization pattern to the nuclear envelope. Colocalizes with ACAA2 in the mitochondria. - Information by UniProt
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参照データベース
- Entrez Gene: 664 Human
- Entrez Gene: 12176 Mouse
- Entrez Gene: 84480 Rat
- Omim: 603293 Human
- SwissProt: Q12983 Human
- SwissProt: O55003 Mouse
- Unigene: 144873 Human
- Unigene: 378890 Mouse
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別名
- BCL2 Adenovirus E1B 19kDa Interacting Protein 3 antibody
- BCL2/adenovirus E1B 19 kDa protein interacting protein 3 antibody
- BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 antibody
see all
画像
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All lanes : Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/1000 dilution
Lane 1 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by RIPA method
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20ug, prepared by 1% SDS hot method
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 21-30,40-60 kDa why is the actual band size different from the predicted?
Exposure time: 156 secondsThe observed band sizes are consistent with publications. Bands around 30 and 60kDa should be monomer and dimer, respectively. Other bands should be different migrating forms (PMID: 15375294, PMID: 36396625, PMID: 9396766, PMID: 36396625, PMID: 33459136 etc.).
1% SDS hot lysis method could help to get stronger monomer forms.The protocol used for this test can be found here
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Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
Lane 2: BNIP3 knockout HAP1 whole cell lysate (40 µg)
Lane 3: SHSY5Y whole cell lysate (40 µg)
Lane 4: A431 whole cell lysate (40 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab109362 observed at 25 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab109362 was shown to recognize BNIP3 in wild-type HAP1 cells as signal was lost at the expected MW in BNIP3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BNIP3 knockout samples were subjected to SDS-PAGE. ab109362 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
All lanes : Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/2000 dilution (purified)
Lane 1 : Rat kidney tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 22 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
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All lanes : Anti-BNIP3 antibody [EPR4034] (ab109362) at 1/1000 dilution (unpurified)
Lane 1 : Jurkat cell lysate treated with etoposide
Lane 2 : Jurkat cell lysate
Lane 3 : SH-SY5Y cell lysate treated with camptothecin
Lane 4 : SH-SY5Y cell lysate
Lane 5 : MCF-7 cell lysate treated with Cocl2
Lysates/proteins at 10 µg per lane.
Predicted band size: 22 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human renal adenocarcinoma tissue labelling BNIP3 with purified ab109362 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [EPR4034] (ab109362)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling BNIP3 with unpurified ab109362 at a 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling BNIP3 with unpurified ab109362 at a 1/100 dilution.
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
参考文献 (77)
ab109362 は 77 報の論文で使用されています。
- Cao Y et al. A mitochondrial SCF-FBXL4 ubiquitin E3 ligase complex degrades BNIP3 and NIX to restrain mitophagy and prevent mitochondrial disease. EMBO J 42:e113033 (2023). PubMed: 36896912
- Yang L et al. Clinical significance and immune landscape of a novel ferroptosis-related prognosis signature in osteosarcoma. BMC Cancer 23:229 (2023). PubMed: 36899330
- Tan Z et al. Transcutaneous Electrical Nerve Stimulation (TENS) Alleviates Brain Ischemic Injury by Regulating Neuronal Oxidative Stress, Pyroptosis, and Mitophagy. Mediators Inflamm 2023:5677865 (2023). PubMed: 37101593
- Elcocks H et al. FBXL4 ubiquitin ligase deficiency promotes mitophagy by elevating NIX levels. EMBO J 42:e112799 (2023). PubMed: 37102372
- Li ZL et al. Activation of HIF-1α C-terminal transactivation domain protects against hypoxia-induced kidney injury through hexokinase 2-mediated mitophagy. Cell Death Dis 14:339 (2023). PubMed: 37225700