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BAX Rabbit Polyclonal Antibody

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Catalog# ER0907

BAX Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

BAX Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within Human BAX aa 1-50 / 192.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 21 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, Daudi cell lysate, Raji cell lysate, Jurkat cell lysate, HepG2, F9, SHSY5Y, human colon carcinoma tissue, mouse kidney tissue.

Conjugation

unconjugated

RRID

AB_3069018

Product Features

Form

Liquid

Concentration

1ug/ul

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:200

  • IHC-P

  • 1:1,000

Target

Function

BAX is a member of the Bcl-2 gene family. Apoptosis regulator BAX promotes apoptosis by binding to and antagonizing the Bcl-2 protein. In healthy mammalian cells, the majority of BAX is found in the cytosol, but upon initiation of apoptotic signaling, Bax undergoes a conformational shift. Upon induction of apoptosis, BAX becomes organelle membrane-associated, and in particular, mitochondrial membrane associated. The expression of BAX is upregulated by the tumor suppressor protein p53, and BAX has been shown to be involved in p53-mediated apoptosis. The p53 protein is a transcription factor that, when activated as part of the cell's response to stress, regulates many downstream target genes, including BAX.

Background References

1. "Elucidation of some Bax conformational changes through crystallization of an antibody-peptide complex." Peyerl F.W., Dai S., Murphy G.A., Crawford F., White J., Marrack P., Kappler J.W. Cell Death Differ. 14:447-452(2006)

2. "BAX activation is initiated at a novel interaction site." Gavathiotis E., Suzuki M., Davis M.L., Pitter K., Bird G.H., Katz S.G., Tu H.C., Kim H., Cheng E.H., Tjandra N., Walensky L.D. Nature 455:1076-1081(2007)

3. "Mutation to Bax beyond the BH3 domain disrupts interactions with pro-survival proteins and promotes apoptosis." Czabotar P.E., Lee E.F., Thompson G.V., Wardak A.Z., Fairlie W.D., Colman P.M. J. Biol. Chem. 286:7123-7131(2010)

Sequence Similarity

Belongs to the Bcl-2 family.

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.

Subcellular Location

Mitochondrion membrane, cytoplasm.

UNIPROT

SwissProt: Q07812 Human

SwissProt: Q07813 Mouse

SwissProt: Q63690 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: MCF7 cell lysate<br />Lane 3: HEK-293 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 21 kDa<br />Observed band size: 21 kDa<br /><br />Exposure time: 1 minute 34 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (ER0907) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: MCF7 cell lysate
    Lane 3: HEK-293 cell lysate

    Lysates/proteins at 15 µg/Lane.

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

    Exposure time: 1 minute 34 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER0907) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: MCF7 cell lysate<br />Lane 3: Daudi cell lysate<br />Lane 4: Raji cell lysate<br />Lane 5: Jurkat cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 21 kDa<br />Observed band size: 21 kDa<br /><br />Exposure time: 29 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (ER0907) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: MCF7 cell lysate
    Lane 3: Daudi cell lysate
    Lane 4: Raji cell lysate
    Lane 5: Jurkat cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 29 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER0907) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/20,000 dilution.<br /><br />Lane 1: MCF7-si NT cell lysate<br />Lane 2: MCF7-si BAX cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 21 kDa<br />Observed band size: 21 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a> was shown to specifically react with BAX in MCF7-si NT cells. Weakened band was observed when MCF7-si BAX sample was tested. MCF7-si NT and MCF7-si BAX samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>, 1/20,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of BAX on different lysates with Rabbit anti-BAX antibody (ER0907) at 1/20,000 dilution.

    Lane 1: MCF7-si NT cell lysate
    Lane 2: MCF7-si BAX cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 3 minutes; ECL: K1802;
    4-20% SDS-PAGE gel.

    ER0907 was shown to specifically react with BAX in MCF7-si NT cells. Weakened band was observed when MCF7-si BAX sample was tested. MCF7-si NT and MCF7-si BAX samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ER0907, 1/20,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of SHSY5Y cells labeling BAX with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta Ⅲ tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 647, <a href="/products/HA1127" style="font-weight: bold;text-decoration: underline;">HA1127</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SHSY5Y cells labeling BAX with Rabbit anti-BAX antibody (ER0907) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-BAX antibody (ER0907) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta Ⅲ tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.

  • ICC staining Bax in HepG2 cells (green). The nuclear counter stain is DAPI (blue).Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining Bax in HepG2 cells (green). The nuclear counter stain is DAPI (blue).Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining Bax in F9 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining Bax in F9 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-BAX antibody (ER0907) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0907) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-BAX antibody (ER0907) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0907) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-BAX antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0907" style="font-weight: bold;text-decoration: underline;">ER0907</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-BAX antibody (ER0907) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0907) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

Filter:
  1. WB
  2. IHC
  3. IF

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