Overview

  • Product nameAnti-Bax antibody [2D2]
    See all Bax primary antibodies
  • Description
    Mouse monoclonal [2D2] to Bax
  • SpecificityThe immunogen amino acid sequence is NOT shared by mouse and rat bax protein.
  • Tested applicationsFlow Cyt, Sandwich ELISA, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cow, Cat, Dog, Chimpanzee, Macaque Monkey

    Does not react with

    Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human Bax aa 3-16 (Cysteine residue).
    Sequence:

    C-GSGEQPRGGGPTSS


    Database link: Q07812

  • Positive control
    • HeLa cells. Hodgkin’s lymphoma.
  • General notes

    This antibody is BSA and azide free

Properties

Applications

Our Abpromise guarantee covers the use of ab77566 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells.
Sandwich ELISA Use a concentration of 0.2 µg/ml. Can be paired for Sandwich ELISA with Rabbit monoclonal [E63] to Bax (ab32503).
WB Use a concentration of 1 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IHC-P Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

Target

  • FunctionAccelerates programmed cell death by binding to, and antagonizing the apoptosis repressor BCL2 or its adenovirus homolog E1B 19k protein. Under stress conditions, undergoes a conformation change that causes translocation to the mitochondrion membrane, leading to the release of cytochrome c that then triggers apoptosis. Promotes activation of CASP3, and thereby apoptosis.
  • Tissue specificityExpressed in a wide variety of tissues. Isoform Psi is found in glial tumors. Isoform Alpha is expressed in spleen, breast, ovary, testis, colon and brain, and at low levels in skin and lung. Isoform Sigma is expressed in spleen, breast, ovary, testis, lung, colon, brain and at low levels in skin. Isoform Alpha and isoform Sigma are expressed in pro-myelocytic leukemia, histiocytic lymphoma, Burkitt's lymphoma, T-cell lymphoma, lymphoblastic leukemia, breast adenocarcinoma, ovary adenocarcinoma, prostate carcinoma, prostate adenocarcinoma, lung carcinoma, epidermoid carcinoma, small cell lung carcinoma and colon adenocarcinoma cell lines.
  • Sequence similaritiesBelongs to the Bcl-2 family.
  • DomainIntact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.
  • Cellular localizationCytoplasm and Mitochondrion membrane. Cytoplasm. Colocalizes with 14-3-3 proteins in the cytoplasm. Under stress conditions, undergoes a conformation change that causes release from JNK-phosphorylated 14-3-3 proteins and translocation to the mitochondrion membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apoptosis regulator BAX antibody
    • Apoptosis regulator BAX cytoplasmic isoform beta antibody
    • Apoptosis regulator BAX membrane isoform alpha antibody
    • BAX antibody
    • Bax isoform psi antibody
    • BAX protein cytoplasmic isoform delta antibody
    • Bax protein cytoplasmic isoform gamma antibody
    • Bax zeta antibody
    • Bax-protein antibody
    • BAX_HUMAN antibody
    • BAXA antibody
    • Bcl-2-like protein 4 antibody
    • BCL2 associated X protein antibody
    • BCL2 associated X protein omega antibody
    • BCL2 associated X protein transcript variant delta2 antibody
    • Bcl2-L-4 antibody
    • BCL2L4 antibody
    • membrane isoform alpha antibody
    see all

Anti-Bax antibody [2D2] images



  • Predicted band size : 21 kDa
    Lane 1 = Bax protein (Tagged) (ab85157) ? 10 ng. Lane 2 = Extract of HeLa cells ? 40 ug. Lane 3 = Extract of HepG2 cells ? 40 ug. Lane 4 = Bax protein (Tagged) (ab85157) ? 10 ng. Lane 5 = Extract of HeLa cells ? 40 ug. Lane 6 = Extract of HepG2 cells ? 40 ug. SDS PAGE performed under reducing conditions (100mM DTT ? Sample heated at 50?C). Primary : Lanes 1-3: Anti Bax antibody (ab77566) at 1 ug/mL. Lanes 4-6: Anti Bax antibody (ab32503) at 1:2000 dilution. Secondary : Lanes 1-3: Goat anti mouse IgG(H&L)-HRP at 1:10000. Lanes 4-6: Goat anti rabbit IgG(H&L)-HRP at 1:10000. Development: ECL with 2 min exposure. Blocking: in 5% Milk + PBS for 3 hours at RT. Primary antibody: in 5% Milk + PBS overnight at 4 C. Secondary antibody: in 5% Milk + PBS for 2 hour at RT. Predicted band size : Bax 21kDa and Bax (Tagged) 49 kDa. Observed band size : Bax 21kDa and Bax (Tagged) 49 kDa
  • Anti-Bax antibody [2D2] (ab77566) at 1 µg/ml + HeLa cell lysate

    Predicted band size : 21 kDa
  • Immunohistochemical analysis of Human Hodgkin's lymphoma tissue, staining Bax with ab77566 at 4 µg/ml.

  • Overlay histogram showing HeLa cells stained with ab77566 (red line). The cells were fixed with 80% methanol (5 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 (ab77566, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Standard Curve for Bax (Analyte: ab85157) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [2D2] to Bax - BSA and Azide free (ab77566) at 0.2ug/ml and Detector Antibody Rabbit monoclonal [E63] to Bax (ab32503) at 0.5ug/ml Concentration of ab32503 may vary from lot to lot; please use this curve as guideline.

Protocols

References for Anti-Bax antibody [2D2] (ab77566)

This product has been referenced in:
  • Arici S  et al. Central nervous system toxicity after acute oral formaldehyde exposure in rabbits: An experimental study. Hum Exp Toxicol N/A:N/A (2014). IHC ; Rabbit . Read more (PubMed: 24501105) »
  • Hsu YT & Youle RJ Nonionic detergents induce dimerization among members of the Bcl-2 family. J Biol Chem 272:13829-34 (1997). WB . Read more (PubMed: 9153240) »

See all 3 Publications for this product

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