Anti-BNIP3抗体[ANa40] (ab10433)

概述

  • 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
  • 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."
  • 经测试应用
    适用于: ICC, ELISA, IP, WB, IHC-P, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant fragment (Residues 1-163 of Human BNIP3).

  • 表位
    The epitope recognized by the antibody resides within amino acids 112-124 of human BNIP3 molecule.
  • 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.
  • 常规说明

    This antibody clone is manufactured by Abcam. 

    Alternative versions available:

    Anti-BNIP3 antibody (Alexa Fluor® 647) [ANa40] (ab196706)

    Anti-BNIP3 antibody (HRP) [ANa40] (ab196707)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

性能

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.

ICC/IF Use at an assay dependent concentration.

靶标

  • 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.
  • 序列相似性
    Belongs to the NIP3 family.
  • 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
  • 数据库链接
  • 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
    • BNIP 3 antibody
    • BNIP3 antibody
    • BNIP3_HUMAN antibody
    • NIP 3 antibody
    • NIP3 antibody
    see all

图片

  • Immunohistochemical 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.
  • Anti-BNIP3 antibody [ANa40] (ab10433) at 1 µg/ml + Human skeletal muscle tissue lysate - total protein (ab29330) at 20 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 21.5 kDa


    Exposure time : 5 seconds
  • 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.

  • 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.

    See Abreview

  • 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.

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) »

See all 59 Publications for this product

客户评价及客户问答

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
Yes - 0.1% triton x-100
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 08 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 04 2016

Application
Western blot
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jun 26 2015

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1126945.

This replacement vial will be shipped Monda...

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I agree this vial of ab10433 you have received is not working as expected. I would be happy to send a free replacement.

So I can arrange the shipment, please provide the PO number or Abcam order reference number associated with the purchase o...

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According to our records, ab10433 was proving difficult to use in WB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details to resolve this case.

As we...

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Thank you for contacting us.

I am sorry to hear that the antibody is not working as before.

As we discussed over the phone, please let me know if a 1/500 dilution helped to improve the results. If not, I'd be happy to offer either ...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Fibroblasts)
Specification
Fibroblasts
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Dr. Sharon Sneddon

Verified customer

提交于 Nov 30 2011

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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