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BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]

Usd: 385

Specification

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Catalog# ET7110-88

BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human BAT3 aa 1,083-1,132 / 1,132.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 119 kDa

Positive Control

293T cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, 293 cell lysate, HepG2 cell lysate, Rat brain tissue lysate, A549, HepG2, rat testis tissue, human lung tissue, human lung carcinoma tissue, SW620.

Conjugation

unconjugated

Clone Number

JE54-90

RRID

AB_3071047

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

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:100

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

Target

Function

ATP-independent molecular chaperone preventing the aggregation of misfolded and hydrophobic patches-containing proteins. Functions as part of a cytosolic protein quality control complex, the BAG6/BAT3 complex, which maintains these client proteins in a soluble state and participates to their proper delivery to the endoplasmic reticulum or alternatively can promote their sorting to the proteasome where they undergo degradation. The BAG6/BAT3 complex is involved in the post-translational delivery of tail-anchored/type II transmembrane proteins to the endoplasmic reticulum membrane. Recruited to ribosomes, it interacts with the transmembrane region of newly synthesized tail-anchored proteins and together with SGTA and ASNA1 mediates their delivery to the endoplasmic reticulum. Client proteins that cannot be properly delivered to the endoplasmic reticulum are ubiquitinated by RNF126, an E3 ubiquitin-protein ligase associated with BAG6 and are sorted to the proteasome. SGTA which prevents the recruitment of RNF126 to BAG6 may negatively regulate the ubiquitination and the proteasomal degradation of client proteins.

Background References

1. Mock JY. et. al. Structural basis for regulation of the nucleo-cytoplasmic distribution of Bag6 by TRC35. Proc Natl Acad Sci U S A. 2017 Oct.

2. Takahashi T. et. al. Cytoplasmic control of Rab family small GTPases through BAG6. EMBO Rep. 2019 Apr.

Tissue Specificity

Expressed by immature dendritic cells (at protein level).

Post-translational Modification

Ricin can induce a cleavage by the caspase CASP3. The released C-terminal peptide induces apoptosis.; (Microbial infection) In case of infection by L.pneumophila, ubiquitinated by the SCF(LegU1) complex.

Subcellular Location

Extracellular exosome, cytosol, nucleus.

UNIPROT

SwissProt: P46379 Human

SwissProt: Q9Z1R2 Mouse

SwissProt: Q6MG49 Rat

Synonyms

2410045D21Rik antibody

AA408914 antibody

BAG 6 antibody

BAG family molecular chaperone regulator 6 antibody

BAG-6 antibody

BAG6 antibody

BAG6_HUMAN antibody

BAT 3 antibody

BAT3 antibody

BCL2-associated athanogene 6 antibody

Expand

Images

  • Western blot analysis of BAT3 on different lysates with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate (20 µg/Lane)<br />Lane 2: Mouse testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 119 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 6 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</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 BAT3 on different lysates with Rabbit anti-BAT3 antibody (ET7110-88) at 1/1,000 dilution.

    Lane 1: 293T cell lysate (20 µg/Lane)
    Lane 2: Mouse testis tissue lysate (40 µg/Lane)

    Predicted band size: 119 kDa
    Observed band size: 150 kDa

    Exposure time: 6 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 (ET7110-88) 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 BAT3 on different lysates with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-BAT3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 119 kDa<br />Observed band size: 160 kDa<br /><br />Exposure time: 60 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/1,000 dilution was used in K1803 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 BAT3 on different lysates with Rabbit anti-BAT3 antibody (ET7110-88) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-BAT3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 119 kDa
    Observed band size: 160 kDa

    Exposure time: 60 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 (ET7110-88) at 1/1,000 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.

  • Western blot analysis of BAT3 on different lysates with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/500 dilution.<br /><br />Lane 1: Rat testis tissue lysate<br />Lane 2: 293 cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: Rat brain tissue lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 119 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/500 dilution was used in 5% NFDM/TBST 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 1:5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of BAT3 on different lysates with Rabbit anti-BAT3 antibody (ET7110-88) at 1/500 dilution.

    Lane 1: Rat testis tissue lysate
    Lane 2: 293 cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: Rat brain tissue lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 119 kDa
    Observed band size: 150 kDa

    Exposure time: 2 minutes;

    6% 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 (ET7110-88) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of A549 cells labeling BAT3 with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 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-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of A549 cells labeling BAT3 with Rabbit anti-BAT3 antibody (ET7110-88) at 1/50 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-BAT3 antibody (ET7110-88) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of HepG2 cells labeling BAT3 with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 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-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling BAT3 with Rabbit anti-BAT3 antibody (ET7110-88) at 1/50 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-BAT3 antibody (ET7110-88) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/200 dilution for 0.5 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 testis tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) at 1/200 dilution for 0.5 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 lung tissue with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/200 dilution for 0.5 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 lung tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) at 1/200 dilution for 0.5 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 lung carcinoma tissue with Rabbit anti-BAT3 antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>) at 1/50 dilution for 0.5 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 lung carcinoma tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) at 1/50 dilution for 0.5 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 BAT3 was done on SW620 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7110-88" style="font-weight: bold;text-decoration: underline;">ET7110-88</a>, 1/50) (red). 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; black).

    Flow cytometric analysis of BAT3 was done on SW620 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-88, 1/50) (red). 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; black).

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