Overview

  • Product name
    Anti-Bax antibody [E63]
    See all Bax primary antibodies
  • Description
    Rabbit monoclonal [E63] to Bax
  • Specificity
    Expression levels of BAX protein vary with sample type. Induction may be required if endogenous expression is low.
  • Tested applications
    Suitable for: IHC-P, WB, IP, Sandwich ELISAmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Chinese hamster
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide within Human Bax aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: HeLa cell lysate. IHC-P: Human lymph node and rat kidney tissues.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

Our Abpromise guarantee covers the use of ab32503 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
IHC-P 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/10000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IP 1/100.
Sandwich ELISA Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Mouse monoclonal [2D2] to Bax (ab77566).
  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Accelerates 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 specificity
      Expressed 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 similarities
      Belongs to the Bcl-2 family.
    • Domain
      Intact 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 localization
      Cytoplasm 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
      • BAX antibody
      • Bax-protein antibody
      • BAX_HUMAN antibody
      • BAXA antibody
      • Baxdelta2G9 antibody
      • Baxdelta2G9omega antibody
      • Baxdelta2omega 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

    Images



    • Predicted band size : 21 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Bax knockout HAP1 cell lysate (20 µg)

      Lanes 1 - 2: Merged signal (red and green). Green - ab32503 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab32503 was shown to recognize Bax when Bax knockout samples were used, along with additional cross-reactive bands. Wild-type and Bax knockout samples were subjected to SDS-PAGE. ab32503 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • IHC image of ab32503 staining Bax in rat kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab32503, 1:250 dilution, 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. No primary antibody was used in the secondary only control (shown on the inset).
      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.



    • Predicted band size : 21 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Bax knockout HAP1 cell lysate (20 µg)

      Lanes 1 - 2: Merged signal (red and green). Green - ab32503 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa or ab18058, observed at 130 kDa.

      This western blot image is a comparison between ab32503 and a competitor's top cited rabbit polyclonal antibody.



    • 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 50oC). 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 [E63] (ab32503) at 1/2000 dilution + HeLa cell lysate

      Secondary
      Goat anti-rabbit HRP antibody
      Developed using the ECL technique

      Predicted band size : 21 kDa
    • Immunohistochemical analysis of paraffin-embedded human lymph node using anti-Bax Rabbit Monoclonal Antibody (ab32503) at 1/250 dilution.

    • 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

    This product has been referenced in:
    • Sun Y  et al. Therapeutic effect of apocynin through antioxidant activity and suppression of apoptosis and inflammation after spinal cord injury. Exp Ther Med 13:952-960 (2017). WB ; Rat . Read more (PubMed: 28450925) »
    • Maunder HE  et al. Enhancing titres of therapeutic viral vectors using the transgene repression in vector production (TRiP) system. Nat Commun 8:14834 (2017). WB . Read more (PubMed: 28345582) »

    See all 116 Publications for this product

    Customer reviews and Q&As

    Filter by Application

    Filter by Species

    Filter by Ratings

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Injured spinal cord)
    Antigen retrieval step
    None
    Permeabilization
    Yes - Triton-X100
    Specification
    Injured spinal cord
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 26°C
    Fixative
    Paraformaldehyde
    Username

    清隆 新井

    Verified customer

    Submitted Sep 19 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (Tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer ph6.0
    Permeabilization
    No
    Specification
    Tonsil
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 26°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Sep 01 2016

    Application
    Western blot
    Sample
    Chinese Hamster Cell lysate - whole cell (CHO cell lines)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    5 µg
    Specification
    CHO cell lines
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 11 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (primary)
    Specification
    primary
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Methanol
    Username

    Abcam user community

    Verified customer

    Submitted Apr 20 2015

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Mouse Cell lysate - whole cell (MEF)
    Specification
    MEF
    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

    Submitted Jul 14 2014

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Rat Cell lysate - whole cell (h9c2)
    Specification
    h9c2
    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

    Submitted Jul 14 2014

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (HEK293)
    Specification
    HEK293
    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

    Submitted Jul 14 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Sample
    Mouse Cell (MEF)
    Specification
    MEF
    Permeabilization
    Yes - 1% TRITON-X-100
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 14 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Sample
    Rat Cell (h9c2)
    Specification
    h9c2
    Permeabilization
    Yes - 1% TRITON-X-100
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 14 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Sample
    Human Cell (HEK293)
    Specification
    HEK293
    Permeabilization
    Yes - 1% TRITON-X-100
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 08 2014

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