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BAP31 Recombinant Rabbit Monoclonal Antibody [JG37-81]

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Specification

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Catalog# ET7108-54

BAP31 Recombinant Rabbit Monoclonal Antibody [JG37-81]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

BAP31 Recombinant Rabbit Monoclonal Antibody [JG37-81]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human BAP31 aa 81-220 / 246.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 28 kDa

Positive Control

A431 cell lysate, SK-Br-3 cell lysate, A431, A549, PC-3M, mouse testis tissue, human testis tissue, Hela.

Conjugation

unconjugated

Clone Number

JG37-81

RRID

AB_3070852

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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

BAP31, a human Bcl-2-interacting protein, is an integral membrane protein that is a component of a protein complex in the endoplasmic reticulum. This protein complex mechanically bridges an apoptosis-initiating caspase, like procaspase-8, with the anti-apoptotic regulator Bcl-2 or Bcl-XL. The cytosolic domain of BAP31 contains two identical caspase recognition sites, which are preferentially cleaved by initiator caspases, including caspase 8. Cleavage of BAP31 during apoptosis generates a p20 fragment, which remains integrated in the membrane and, when expressed ectopically, is a potent inducer of cell death. BAP31 cleavage is important for manifesting cytoplasmic apoptotic events associated with membrane fragmentation and in the cross talk between mitochondria and the endoplasmic reticulum during Fas- mediated apoptosis. The BAP31 gene is ubiquitously expressed in murine tissues and is located on the X chromosome in both mouse and human.

Background References

1. Annaert W G et al. Export of cellubrevin from the endoplasmic reticulum is controlled by BAP31. J Cell Biol 139:1397-1410 (1997).

2. Paquet M E et al. Bap29/31 influences the intracellular traffic of MHC class I molecules. J Immunol 172:7548-7555 (2004).

Sequence Similarity

Belongs to the BCAP29/BCAP31 family.

Tissue Specificity

Ubiquitous. Highly expressed in neurons and discrete endocrine cells.

Post-translational Modification

Cleaved by CASP8 and other caspases.

Subcellular Location

Endoplasmic reticulum membrane.

UNIPROT

SwissProt: P51572 Human

SwissProt: Q61335 Mouse

Synonyms

6C6 AG antibody

6C6 AG tumor associated antigen antibody

6C6-AG tumor-associated antigen antibody

6C6AG antibody

6C6AG tumor associated antigen antibody

Accessory protein BAP 31 antibody

Accessory protein BAP31 antibody

B cell receptor associated protein 31 antibody

B-cell receptor-associated protein 31 antibody

BA31 antibody

Expand

Images

  • Western blot analysis of BAP31 on different lysates with Rabbit anti-BAP31 antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate<br />Lane 2: SK-Br-3 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 28 kDa<br />Observed band size: 28 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/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</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 BAP31 on different lysates with Rabbit anti-BAP31 antibody (ET7108-54) at 1/1,000 dilution.

    Lane 1: A431 cell lysate
    Lane 2: SK-Br-3 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 28 kDa
    Observed band size: 28 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 (ET7108-54) 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.

  • ICC staining of BAP31 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of BAP31 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-54, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of BAP31 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of BAP31 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-54, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of BAP31 in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of BAP31 in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7108-54, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-BAP31 antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</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/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</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 testis tissue with Rabbit anti-BAP31 antibody (ET7108-54) 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 (ET7108-54) 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.

  • Flow cytometric analysis of BAP31 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7108-54" style="font-weight: bold;text-decoration: underline;">ET7108-54</a>, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

    Flow cytometric analysis of BAP31 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7108-54, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).

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