Overview

  • Product name

    Anti-Bax antibody [EPR18283]
    See all Bax primary antibodies
  • Description

    Rabbit monoclonal [EPR18283] 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: IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Bax aa 1-100. The exact sequence is proprietary.
    Database link: Q07812

  • Positive control

    • WB: Human fetal liver, fetal heart, fetal kidney, fetal spleen and fetal lung lysates; Human thymus, cerebellum and lymph node lysates; HeLa, MCF7, HuT-78, Ramos, HEK-293, Raji, L-929, C2C12, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, kidney and spleen lysates. IP: HeLa whole cell lysate.
  • General notes

     

     

    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 ab182733 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
IP 1/50.
WB 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

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

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: BAX knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HT29 whole cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab182733 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab182733 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. Ab182733 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal spleen lysate
    Lane 5 : Human fetal lung lysate
    Lane 6 : Human thymus lysate
    Lane 7 : Human cerebellum lysate
    Lane 8 : Human lymph node lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 21 kDa
    Observed band size: 21 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2,3 and 4: 8 seconds; Lane 5 and 6: 1 minute; Lane 7 and 8: 15 seconds.

  • All lanes : Anti-Bax antibody [EPR18283] (ab182733) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 3 : HuT-78 (Human Sezary syndrome cell line) whole cell lysate
    Lane 4 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 6 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 7 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 8 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
    Lane 9 : C2C12 (Mouse myoblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 21 kDa
    Observed band size: 21 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Jurkat doesn't express Bax. (PMID: 9583678).

  • All lanes : Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : Rat spleen lysate
    Lane 8 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 9 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 10 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 11 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 21 kDa
    Observed band size: 21 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-7: 30 seconds; Lane 8-11: 1 second.

     

  • Bax was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182733 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab182733 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab182733 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab182733 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

  • All lanes : Anti-Bax antibody [EPR18283] (ab182733) at 1/2000 dilution

    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
    Lane 8 : Rat brain. Whole tissue lysate
    Lane 9 : 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 58 s

References

This product has been referenced in:

  • Zhao L  et al. Saikosaponin D inhibits proliferation of human osteosarcoma cells via the p53 signaling pathway. Exp Ther Med 17:488-494 (2019). Read more (PubMed: 30651826) »
  • E L  et al. MicroRNA-144 attenuates cardiac ischemia/reperfusion injury by targeting FOXO1. Exp Ther Med 17:2152-2160 (2019). Read more (PubMed: 30783480) »
See all 37 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
HEK293
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 24 2018

Application
Western blot
Sample
Rat Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 24 2018

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

Abcam user community

Verified customer

Submitted Dec 24 2018

Application
Immunocytochemistry
Sample
Rat Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 28 2018

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Non-reduced Denaturing (12%)
Loading amount
30 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Jun 30 2017

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