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Bak Recombinant Rabbit Monoclonal Antibody [SU32-07]

Usd: 385

Specification

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Catalog# ET1608-21

Bak Recombinant Rabbit Monoclonal Antibody [SU32-07]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

Bak Recombinant Rabbit Monoclonal Antibody [SU32-07]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Bak aa 1-50 / 211.

Species Reactivity

Human

Validated Applications

WB, IF-Tissue, IHC-P, IP

Molecular Weight

Predicted band size: 23 kDa

Positive Control

HeLa cell lysate, AGS cell lysate, HEK-293 cell lysate, human colon tissue, human stomach carcinoma tissue, human small intestine tissue.

Conjugation

unconjugated

Clone Number

SU32-07

RRID

AB_2940739

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • 1:1,000-1:2,000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:500-1:1,000

  • IP

  • 1-2μg/sample

Target

Function

The protein encoded by this gene belongs to the BCL2 protein family. BCL2 family members form oligomers or heterodimers and act as anti- or pro-apoptotic regulators that are involved in a wide variety of cellular activities. This protein localizes to mitochondria, and functions to induce apoptosis. It interacts with and accelerates the opening of the mitochondrial voltage-dependent anion channel, which leads to a loss in membrane potential and the release of cytochrome c. This protein also interacts with the tumor suppressor P53 after exposure to cell stress.

Background References

1. Holloway A et al. Resistance to UV-induced apoptosis by -HPV5 E6 involves targeting of activated BAK for proteolysis by recruitment of the HERC1 ubiquitin ligase. Int J Cancer 136:2831-43 (2015).

2. Wang B et al. Role of Ku70 in deubiquitination of Mcl-1 and suppression of apoptosis. Cell Death Differ 21:1160-9 (2014).

Sequence Similarity

Belongs to the Bcl-2 family.

Tissue Specificity

Expressed in a wide variety of tissues, with highest levels in the heart and skeletal muscle.

Subcellular Location

Mitochondrion membrane.

UNIPROT

SwissProt: Q16611 Human

Synonyms

Apoptosis regulator BAK antibody

BAK antibody

BAK like antibody

Bak NT antibody

BAK_HUMAN antibody

Bak1 antibody

Bcl 2 homologous antagonist/killer antibody

Bcl 2 like 7 protein antibody

Bcl-2 homologous antagonist/killer antibody

Bcl-2-like protein 7 antibody

Expand

Images

  • Western blot analysis of Bak on different lysates with Rabbit anti-Bak antibody (<a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: AGS cell lysate<br />Lane 3: HEK-293 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 23 kDa<br />Observed band size: 23 kDa<br /><br />Exposure time: 3 minutes; 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/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/1,000 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 Bak on different lysates with Rabbit anti-Bak antibody (ET1608-21) at 1/1,000 dilution.

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

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa

    Exposure time: 3 minutes; 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 (ET1608-21) at 1/1,000 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 Bak on different lysates with Rabbit anti-Bak antibody (<a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Bak KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 23 kDa<br />Observed band size: 23 kDa<br /><br />Exposure time: 120 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/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/2,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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 Bak on different lysates with Rabbit anti-Bak antibody (ET1608-21) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Bak KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa

    Exposure time: 120 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 (ET1608-21) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Bak antibody (<a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</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/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</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 tissue with Rabbit anti-Bak antibody (ET1608-21) 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 (ET1608-21) 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 stomach carcinoma tissue with Rabbit anti-Bak antibody (<a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/500 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/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/500 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 stomach carcinoma tissue with Rabbit anti-Bak antibody (ET1608-21) at 1/500 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 (ET1608-21) at 1/500 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 small intestine tissue with Rabbit anti-Bak antibody (<a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/500 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/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a>) at 1/500 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 small intestine tissue with Rabbit anti-Bak antibody (ET1608-21) at 1/500 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 (ET1608-21) at 1/500 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.

  • Bak was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1608-21" style="font-weight: bold;text-decoration: underline;">ET1608-21</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Bak was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1608-21 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1608-21 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: ET1608-21 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ET1608-21 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute; ECL: K1801

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Citation