- 数据表
- 具体的参考文献 (56)
- 实验方案
概述
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产品名称Anti-BNIP3抗体[ANa40]
参阅全部 BNIP3 一抗 -
描述小鼠单克隆抗体[ANa40] to BNIP3
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特异性This antibody was originally tested on whole cell lysate from 293T cells transfected with BNIP3. Levels of constitutively expressed BNIP3 are very variable and a good positive control is strongly recommended. Zhang et aln (2008) comment "Very low levels of BNIP3 protein were detected by immunoblot assay of lysates prepared from WT MEFs that were cultured at 20% O2, whereas hypoxia strongly induced expression of BNIP3 protein, which migrated as a 30-kDa monomer and 60-kDa dimer, as previously described. ... The expression of BNIP3 mRNA and protein was significantly reduced in the lungs of HET (as compared with WT) and Cre+ (as compared with Cre-) mice, demonstrating that BNIP3 expression is regulated by HIF-1 under physiological conditions in vivo."
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经测试应用适用于: ICC, ELISA, IP, WB, IHC-P, Flow Cyt, ICC/IFmore details
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种属反应性与反应: Mouse, Rat, Human
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免疫原
Recombinant fragment (Residues 1-163 of Human BNIP3).
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表位The epitope recognized by the antibody resides within amino acids 112-124 of human BNIP3 molecule.
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阳性对照
- This antibody gave a positive signal in human skeletal muscle tissue lysate in western blot and on human kidney formalin-fixed, paraffin-embedded tissue sections in IHC.
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常规说明
BNIP3, formerly NIP3 (nineteen kDa interacting protein-3), is a pro-apoptotic, mitochondrial protein classified in the Bcl 2 family based on limited sequence homology to the Bcl 2 homology 3 (BH3) domain (amino acids 110-118) and C-terminal TM domain. BNIP3 expressed in yeast and mammalian cells interacts with survival promoting proteins Bcl 2, Bcl XL, CED9 and the adenovirus E1B 19K protein. Typically the BH3 domain of pro-apoptotic Bcl-2 homologues mediates Bcl 2/Bcl XL heterodimerization and confers pro-apoptotic activity. BNIP3 represents a subfamily of Bcl 2 related proteins, which functions without a typical BH3 domain to regulate apoptosis from both mitochondrial and nonmitochondrial sites by selective Bcl 2/Bcl XL interactions. The N-terminus (residues 1-49) and the C-terminus TM domain of BNIP3 are critical for Bcl 2 heterodimerization, and either region is sufficient for Bcl XL interaction. The TM domain of BNIP3 is critical for homodimerization, pro-apoptotic function, and mitochondrial targeting. BNIP3 contains PEST sequences suggesting that the protein may be susceptible to rapid degradation by proteases. PEST sequences commonly contain high local concentrations of amino acids P, E, S, T, and D flanked by charged amino acids and these are abundantly present in NIP3. Thus, the posttranslational control of BNIP3 expression through rapid protein degradation may constitute a mechanism for regulating the intracellular levels of a potentially lethal protein. Homologues of BNIP3 sharing both structural and functional similarity have been identified in mammals: Nix (also called BNIP3L/BNIP3alpha\/B5) and, in C. elegans, ceBNIP3. The TM domain of BNIP3 and Nix share 80% identity. Endogenous BNIP3 is loosely associated with mitochondrial membrane in normal tissue but fully integrates into the mitochondrial outer membrane with the N-terminus in the cytoplasm and the C-terminus in the membrane during induction of cell death. This is accompanied by rapid and profound mitochondrial dysfunction characterized by opening of the mitochondrial permeability transition (PT) pore, proton electrochemical gradient suppression, and increased reactive oxygen species production. BNIP3 has been reported to localize to the nuclear envelope when co-expressed with E1B 19K. Endogenous BNIP3 protein is abundant in murine and human skeletal muscle and is not detectable in lysates of all other nonskeletal muscle-bearing tissues and many cell lines, including myoblasts and differentiated myocytes.
Alternative versions available:
Anti-BNIP3 antibody (Alexa Fluor® 647) [ANa40] (ab196706)
Anti-BNIP3 antibody (HRP) [ANa40] (ab196707)
性能
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形式Liquid
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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存储溶液pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: 0.0268% PBS
Some batches contain 0.4M arginine. Please contact Scientific Support for further information. -
Concentration information loading... -
纯度Protein G purified
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Primary antibody说明BNIP3, formerly NIP3 (nineteen kDa interacting protein-3), is a pro-apoptotic, mitochondrial protein classified in the Bcl 2 family based on limited sequence homology to the Bcl 2 homology 3 (BH3) domain (amino acids 110-118) and C-terminal TM domain. BNIP3 expressed in yeast and mammalian cells interacts with survival promoting proteins Bcl 2, Bcl XL, CED9 and the adenovirus E1B 19K protein. Typically the BH3 domain of pro-apoptotic Bcl-2 homologues mediates Bcl 2/Bcl XL heterodimerization and confers pro-apoptotic activity. BNIP3 represents a subfamily of Bcl 2 related proteins, which functions without a typical BH3 domain to regulate apoptosis from both mitochondrial and nonmitochondrial sites by selective Bcl 2/Bcl XL interactions. The N-terminus (residues 1-49) and the C-terminus TM domain of BNIP3 are critical for Bcl 2 heterodimerization, and either region is sufficient for Bcl XL interaction. The TM domain of BNIP3 is critical for homodimerization, pro-apoptotic function, and mitochondrial targeting. BNIP3 contains PEST sequences suggesting that the protein may be susceptible to rapid degradation by proteases. PEST sequences commonly contain high local concentrations of amino acids P, E, S, T, and D flanked by charged amino acids and these are abundantly present in NIP3. Thus, the posttranslational control of BNIP3 expression through rapid protein degradation may constitute a mechanism for regulating the intracellular levels of a potentially lethal protein. Homologues of BNIP3 sharing both structural and functional similarity have been identified in mammals: Nix (also called BNIP3L/BNIP3alpha\/B5) and, in C. elegans, ceBNIP3. The TM domain of BNIP3 and Nix share 80% identity. Endogenous BNIP3 is loosely associated with mitochondrial membrane in normal tissue but fully integrates into the mitochondrial outer membrane with the N-terminus in the cytoplasm and the C-terminus in the membrane during induction of cell death. This is accompanied by rapid and profound mitochondrial dysfunction characterized by opening of the mitochondrial permeability transition (PT) pore, proton electrochemical gradient suppression, and increased reactive oxygen species production. BNIP3 has been reported to localize to the nuclear envelope when co-expressed with E1B 19K. Endogenous BNIP3 protein is abundant in murine and human skeletal muscle and is not detectable in lysates of all other nonskeletal muscle-bearing tissues and many cell lines, including myoblasts and differentiated myocytes.
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克隆单克隆
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克隆编号ANa40
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骨髓瘤unknown
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同种型IgG2b
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轻链类型kappa
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研究领域
相关产品
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
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Related Products
应用
Our Abpromise guarantee covers the use of ab10433 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC | Use at an assay dependent concentration. | |
| ELISA | Use at an assay dependent concentration. | |
| IP | Use at an assay dependent concentration. | |
| WB | Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 21.5 kDa). | |
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| Flow Cyt | Use 0.1-1µg for 106 cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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| ICC/IF | Use at an assay dependent concentration. |
靶标
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功能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.
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序列相似性Belongs to the NIP3 family.
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细胞定位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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数据库链接
- 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
see all -
别名
- 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
Anti-BNIP3 antibody [ANa40] 图像
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [ANa40] (ab10433)](http://www.abcam.cn/ps/products/10/ab10433/Images/BNIP3-Primary-antibodies-ab10433-10.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [ANa40] (ab10433)Image from Jin T et al., J Transl Med. 2012 Mar 30;10:64 Fig 1.; doi:10.1186/1479-5876-10-64; 30 March 2012, Journal of Translational Medicine 2012, 10:64Immunohistochemical analysis of Human laryngeal squamous cell carcinoma tissue, staining BNIP3 with ab10433.
Antigen retrieval was performed by heat mediation in citrate buffer. The sections were incubated with primary antibody (1/100) overnight at 4°C in a humidified chamber. Staining was visualized using DAB, followed by hematoxylin nuclear counterstaining. -
![Western blot - Anti-BNIP3 antibody [ANa40] (ab10433)](http://www.abcam.cn/ps/products/10/ab10433/Images/BNIP3-Primary-antibodies-ab10433-11.jpg)
Western blot - Anti-BNIP3 antibody [ANa40] (ab10433)Anti-BNIP3 antibody [ANa40] (ab10433) at 5 µg/ml + Human skeletal muscle tissue lysate - total protein (ab29330) at 20 µg
Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 21.5 kDa
Observed band size : 30 kDa (why is the actual band size different from the predicted?)
Additional bands at : 135 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [ANa40] (ab10433)](http://www.abcam.cn/ps/products/10/ab10433/Images/BNIP3-Primary-antibodies-ab10433-9.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BNIP3 antibody [ANa40] (ab10433)IHC image of ab10433 staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10433, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
![Immunocytochemistry/ Immunofluorescence - Anti-BNIP3 antibody [ANa40] (ab10433)](http://www.abcam.cn/ps/products/10/ab10433/Images/BNIP3-Primary-antibodies-ab10433-6.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-BNIP3 antibody [ANa40] (ab10433)This image is courtesy of an Abreview submitted by Sharon Sneddon.ab10433 staining BNIP3 in Human fibroblasts by Immunocytochemistry/ Immunofluorescence. Cells were fixed in 2% paraformaldehyde for 30 minutes at room temperature and cells were treated with 50 mM NH4Cl in PBS for 10 minutes to quench free aldehyde groups after permeabilisation. Cells were then permeabilized in 0.1% Triton X-100 prior to blocking in 1% BSA for 1 hour at room temperature. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at room temperature. The secondary antibody was Alexa Fluor®555-conjugated goat anti-mouse polyclonal, diluted 1/1000. -
![Flow Cytometry - Anti-BNIP3 antibody [ANa40] (ab10433)](http://www.abcam.cn/ps/products/10/ab10433/Images/ab10433-17999-ab10433FC.jpg)
Flow Cytometry - Anti-BNIP3 antibody [ANa40] (ab10433)Overlay histogram showing HepG2 cells stained with ab10433 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab10433, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Anti-BNIP3 antibody [ANa40] (ab10433)参考文献
This product has been referenced in:
- Ono M et al. Enhanced snoMEN Vectors Facilitate Establishment of GFP-HIF-1a Protein Replacement Human Cell Lines. PLoS One 11:e0154759 (2016). ICC/IF ; Human . Read more (PubMed: 27128805) »
- Sun Y et al. A novel TP53 pathway influences the HGS-mediated exosome formation in colorectal cancer. Sci Rep 6:28083 (2016). WB ; Human . Read more (PubMed: 27312428) »
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