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BAX Recombinant Rabbit Monoclonal Antibody [SZ3-07]

Usd: 385

Specification

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Catalog# ET1603-34

BAX Recombinant Rabbit Monoclonal Antibody [SZ3-07]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Goat

Predicted species support after-sales service
Conjugation

  • unconjugated

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

Overview

Product Name

BAX Recombinant Rabbit Monoclonal Antibody [SZ3-07]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

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

Species Reactivity

Human, Mouse, Rat (Predicted: Goat)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 21 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, bEnd.3 cell lysate, C2C12 cell lysate, PC-12 cell lysate, C6 cell lysate, C2C12, HeLa.

Conjugation

unconjugated

Clone Number

SZ3-07

RRID

AB_3069679

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:20,000-1:50,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

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. BAX is believed to interact with, and induce the opening of the mitochondrial voltage-dependent anion channel, VDAC. Alternatively, growing evidence also suggests that activated BAX and/or Bak form an oligomeric pore, MAC in the MOM (mitochondrial outer membrane). This results in the release of cytochrome c and other pro-apoptotic factors from the mitochondria, often referred to as mitochondrial outer membrane permeabilization, leading to activation of caspases. This defines a direct role for BAX in mitochondrial outer membrane permeabilization. BAX activation is stimulated by various abiotic factors, including heat, hydrogen peroxide, low or high pH, and mitochondrial membrane remodeling. In addition, it can become activated by binding BCL-2, as well as non-BCL-2 proteins such as p53 and Bif-1. Conversely, BAX can become inactivated by interacting with VDAC2, Pin1, and IBRDC2.

Background References

1. He G et al. Gadd45b prevents autophagy and apoptosis against rat cerebral neuron oxygen-glucose deprivation/reperfusion injury. Apoptosis 21:390-403 (2016).

2. Chen B et al. Inhibition of miR-29c promotes proliferation, and inhibits apoptosis and differentiation in P19 embryonic carcinoma cells. Mol Med Rep 13:2527-35 (2016).

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/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>) at 1/20,000 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 />Lane 4: bEnd.3 cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: PC-12 cell lysate<br />Lane 7: C6 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: 30 seconds; ECL: K1801;<br /><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/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>) at 1/20,000 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 (ET1603-34) at 1/20,000 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
    Lane 4: bEnd.3 cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: PC-12 cell lysate
    Lane 7: C6 cell lysate

    Lysates/proteins at 15 µg/Lane.

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

    Exposure time: 30 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 (ET1603-34) at 1/20,000 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.

  • <span style="font-weight: bold;">☑ Knockout (KO)</span><br /><br />All lanes: Western blot analysis of Bax with anti-Bax antibody [SZ3-07] (<a href="/products/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>) at 1/1,000 dilution.<br />Lane 1/2: Wild-type Hela whole cell lysate (20 µg).<br />Lane 3/4: Bax knockout Hela whole cell lysate (20 µg).<br /><br /><a href="/products/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a> was shown to specifically react with Bax in wild-type Hela cells. No band was observed when Bax knockout samples were tested. Wild-type and Bax knockout 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/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>, 1/1,000) and Loading control antibody (Rabbit anti-β-actin, <a href="/products/R1207-1" style="font-weight: bold;text-decoration: underline;">R1207-1</a>, 1/1,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Knockout (KO)

    All lanes: Western blot analysis of Bax with anti-Bax antibody [SZ3-07] (ET1603-34) at 1/1,000 dilution.
    Lane 1/2: Wild-type Hela whole cell lysate (20 µg).
    Lane 3/4: Bax knockout Hela whole cell lysate (20 µg).

    ET1603-34 was shown to specifically react with Bax in wild-type Hela cells. No band was observed when Bax knockout samples were tested. Wild-type and Bax knockout 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 (ET1603-34, 1/1,000) and Loading control antibody (Rabbit anti-β-actin, R1207-1, 1/1,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of C2C12 cells labeling BAX with Rabbit anti-BAX antibody (<a href="/products/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BAX antibody (<a href="/products/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>) at 1/100 dilution in 1% BSA in PBST 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. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of C2C12 cells labeling BAX with Rabbit anti-BAX antibody (ET1603-34) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BAX antibody (ET1603-34) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling BAX.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1603-34" style="font-weight: bold;text-decoration: underline;">ET1603-34</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling BAX.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1603-34, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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

Citation

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