BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]
Catalog# ET7110-88
BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]
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WB
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IF-Cell
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IHC-P
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FC
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Human
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Mouse
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Rat
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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
Target 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
Reactivity Data
| WB | IF-Cell | IHC-P | FC | |
|---|---|---|---|---|
| Human |
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| Mouse |
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| Rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
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
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WB
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1:1,000-1:2,000
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IF-Cell
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1:50-1:100
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IHC-P
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1:50-1:200
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FC
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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.
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
ExpandImages
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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. -
☑ 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. -
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 (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 (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 (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 (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 (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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