Overview

  • Product name
    Anti-Bax antibody [E63]
    See all Bax primary antibodies
  • Description
    Rabbit monoclonal [E63] to Bax
  • Host species
    Rabbit
  • 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.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • WB: HeLa cell lysate. IHC-P: Human lymph node and rat kidney tissues.
  • General notes

    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

    This product is a recombinant rabbit monoclonal antibody.

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
      see all
    • 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

    • Immunoprecipitation - Anti-Bax antibody [E63] (ab32503)
      Immunoprecipitation - Anti-Bax antibody [E63] (ab32503)

      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg
      Lane 2 (+): HeLa whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32503 in HeLa whole cell lysate

      Purified ab32503 immunoprecipitating Bax in HeLa lysates. For western blotting, the primary antibody used was purified ab32503 at 1/1000 dilution. Ab131366 VeriBlot for IP (HRP) was used as the secondary antibody at 1/1000 dilution. Capture antibody was used at a 1/20 dilution. Blocking and diluting buffer used was 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)

      Purified ab32503 staining Bax in Human lung carcinoma tissue section by immmunohistochemistry (IHC-P- Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraffin and heat mediated antigen retrieval was performed using EDTA buffer (pH 9.0). Samples were incubated with primary antibody at 1:500 dilution. A goat anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody at 1:500 dilution. Cytoplasmic staining on human lung carcinoma.

    • Western blot - Anti-Bax antibody [E63] (ab32503)
      Western blot - Anti-Bax antibody [E63] (ab32503)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)

      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.

    • Western blot - Anti-Bax antibody [E63] (ab32503)
      Western blot - Anti-Bax antibody [E63] (ab32503)

      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.

    • Western blot - Anti-Bax antibody [E63] (ab32503)
      Western blot - Anti-Bax antibody [E63] (ab32503)

      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.

    • Western blot - Anti-Bax antibody [E63] (ab32503)
      Western blot - Anti-Bax antibody [E63] (ab32503)
      All lanes : Anti-Bax antibody [E63] (ab32503) at 1/2000 dilution (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell). Whole cell lysates
      Lane 2 : Hep G2 (Human hepatocellular carcinoma epithelial cell). Whole cell lysates
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) Whole cell lysates
      Lane 4 : A549 (Human lung carcinoma epithelial cell) Whole cell lysates
      Lane 5 : C2C12 (Mouse myoblasts myoblast) Whole cell lysates
      Lane 6 : C6 (Rat glial tumor glial cell) Whole cell lysates
      Lane 7 : Mouse brain. Whole tissue lysate
      Lanes 8 & 10 : Rat brain. Whole tissue lysate
      Lanes 9 & 11 : Rat Spleen. Whole tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 21 kDa
      Observed band size: 18 kDa (why is the actual band size different from the predicted?)



      Blocking and Diluting buffers: 5% NFDM/TBST

      Exposure time 1~9 lanes 32 s; 10~11 lanes 3 min

      Jurkat is negative reported by PMID: 15528359. Brain is low expressed reported by PMID: 27069530.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] (ab32503)

      Immunohistochemical analysis of paraffin-embedded human lymph node using anti-Bax Rabbit Monoclonal Antibody (ab32503) at 1/250 dilution.

    • Sandwich ELISA - Anti-Bax antibody [E63] (ab32503)
      Sandwich ELISA - Anti-Bax antibody [E63] (ab32503)
      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:
    • An MJ  et al. Transcriptome analysis for UVB-induced phototoxicity in mouse retina. Environ Toxicol 33:52-62 (2018). Read more (PubMed: 29044990) »
    • Sun D  et al. bFGF plays a neuroprotective role by suppressing excessive autophagy and apoptosis after transient global cerebral ischemia in rats. Cell Death Dis 9:172 (2018). WB ; Rat . Read more (PubMed: 29416039) »

    See all 177 Publications for this product

    Publishing research using ab32503? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    14 reviews or Q&A

    10
    Votes: Highest First

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Bax [E63] antibody

     Excellent
    Abreviews
    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
    Read More
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    Username

    Abcam user community

    Verified customer

    Submitted Jul 08 2014

    Western blot abreview for Anti-Bax antibody [E63]

     Excellent
    Abreviews
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing (13%)
    Sample
    Human Cell lysate - whole cell (HCT116 cell lysates)
    Specification
    HCT116 cell lysates
    Treatment
    DMSO control (Ctl) and 7,5 ´M CPT-11 (CPT) for 24 hrs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
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    Username

    Mr. Christian Marx

    Verified customer

    Submitted Nov 29 2013

    Western blot abreview for Anti-Bax antibody [E63]

     Excellent
    Abreviews
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing (13%)
    Sample
    Mouse Cell lysate - whole cell (MEF cell lysates)
    Specification
    MEF cell lysates
    Treatment
    DMSO control (Ctl) and 7,5 ´M CPT-11 (CPT) for 24 hrs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    abreview image
    Username

    Mr. Christian Marx

    Verified customer

    Submitted Nov 29 2013

    Western blot abreview for Anti-Bax antibody [E63]

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Muller stem cells mio-m1 total cell lysate)
    Loading amount
    40 µg
    Specification
    Human Muller stem cells mio-m1 total cell lysate
    Gel Running Conditions
    Reduced Denaturing (4-12% gradient gel, sample was denatured at 70C for 10mins)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
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    Username

    Abcam user community

    Verified customer

    Submitted Nov 11 2011

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