Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Bax antibody [EPR18284] (ab182734)

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Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
  • Immunoprecipitation - Anti-Bax antibody [EPR18284] (ab182734)
  • Anti-Bax antibody [EPR18284] (ab182734)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18284] to Bax
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Bax antibody [EPR18284]
    See all Bax primary antibodies

  • Description

    Rabbit monoclonal [EPR18284] 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: WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, MCF7, HuT-78, Ramos, HEK-293, Raji, L-929, C2C12, C6, PC-12 and NIH/3T3 whole cell lysates; Human kidney and spleen lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, kidney and spleen lysates; IP: HeLa whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab182734 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
WB
1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 21 kDa).
IP
1/40.
Notes
WB
1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 21 kDa).
IP
1/40.

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

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : BAX knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



    Lanes 1- 2: Merged signal (red and green). Green - ab182734 observed at 21 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab182734 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : BAX knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : HT29 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 21 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab182734 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab182734 was shown to specifically react with BAX when BAX knockout samples were used. Wild-type and BAX knockout samples were subjected to SDS-PAGE. Ab182734 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 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: 19 kDa why is the actual band size different from the predicted?


    Exposure time: 2 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    Negative control:

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

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 dilution

    Lane 1 : Human kidney lysate
    Lane 2 : Human spleen 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: 19 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    This product reacts with a 15 kD isoform (PMID: 7607685).

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734)

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : Bax knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 21 kDa



    Lanes 1 - 2: Merged signal (red and green). Green - ab182734 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 ab182734 and a competitor's top cited rabbit polyclonal antibody.

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) at 1/1000 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 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 10 : 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: 19 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1, 2 and 3: 3 seconds; Lane 4: 2 seconds; Lane 5: 3 seconds; Lane 6 and 7: 2 seconds; Lane 8, 9 and 10: 1 second.

    This product reacts with a 15 kD isoform (PMID: 7607685).

  • Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    Western blot - Anti-Bax antibody [EPR18284] (ab182734)
    All lanes : Anti-Bax antibody [EPR18284] (ab182734) 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
    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 5% NFDM/TBST

    Exposure time 1~9 lanes 24 s; 10~11 lanes 3 min

  • Immunoprecipitation - Anti-Bax antibody [EPR18284] (ab182734)
    Immunoprecipitation - Anti-Bax antibody [EPR18284] (ab182734)

    Bax was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182734 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab182734 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: ab182734 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab182734 in HeLa whole cell lysate.

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

    Exposure time: 10 seconds.

  • Anti-Bax antibody [EPR18284] (ab182734)
    Anti-Bax antibody [EPR18284] (ab182734)

References (51)

Publishing research using ab182734? Please let us know so that we can cite the reference in this datasheet.

ab182734 has been referenced in 51 publications.

  • Zhou Q  et al. Resveratrol ameliorates neuronal apoptosis and cognitive impairment by activating the SIRT1/RhoA pathway in rats after anesthesia with sevoflurane. Bosn J Basic Med Sci 22:110-117 (2022). PubMed: 34247566
  • Zeng C  et al. Repressing phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma by microRNA-142-3p restrains the progression of hepatocellular carcinoma. Bioengineered 13:1491-1506 (2022). PubMed: 34986757
  • Zhou Q  et al. Safety Profile of Rapamycin Perfluorocarbon Nanoparticles for Preventing Cisplatin-Induced Kidney Injury. Nanomaterials (Basel) 12:N/A (2022). PubMed: 35159680
  • Wang L  et al. Long Noncoding RNA Solute Carrier Family 25 Member 21 Antisense RNA 1 Inhibits Cell Malignant Behaviors and Enhances Radiosensitivity of Gastric Cancer Cells by Upregulating Synuclein Gamma Expression. Tohoku J Exp Med 257:225-239 (2022). PubMed: 35444105
  • Yin X  et al. Dexmedetomidine Combined with Low-Dose Propofol Declines Learning and Memory Impairment and Neural Cell Injury in Developing Rats. Evid Based Complement Alternat Med 2022:9543459 (2022). PubMed: 35685722
View all Publications for this product

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