Bag1 Recombinant Rabbit Monoclonal Antibody [JE63-22]
Catalog# HA721885
Bag1 Recombinant Rabbit Monoclonal Antibody [JE63-22]
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WB
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IHC-P
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IF-Cell
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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
Bag1 Recombinant Rabbit Monoclonal Antibody [JE63-22]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Bag1 aa 111-160 / 345.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, FC
Target Molecular Weight
Predicted band size: 38 kDa
Positive Control
HeLa cell lysate, Jurkat cell lysate, HEK-293 cell lysate, MCF7 cell lysate, RAW264.7 cell lysate, rat testis tissue lysate, mouse testis tissue lysate, human breast cancer tissue, human prostate cancer tissue, HeLa.
Conjugation
unconjugated
Clone Number
JE63-22
RRID
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
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IHC-P
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1:500
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IF-Cell
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1:100
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FC
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1:1,000
Target
Function
Co-chaperone for HSP70 and HSC70 chaperone proteins. Acts as a nucleotide-exchange factor (NEF) promoting the release of ADP from the HSP70 and HSC70 proteins thereby triggering client/substrate protein release. Nucleotide release is mediated via its binding to the nucleotide-binding domain (NBD) of HSPA8/HSC70 where as the substrate release is mediated via its binding to the substrate-binding domain (SBD) of HSPA8/HSC70. Inhibits the pro-apoptotic function of PPP1R15A, and has anti-apoptotic activity. Markedly increases the anti-cell death function of BCL2 induced by various stimuli.
Background References
1. Rauch J.N., Zuiderweg E.R., Gestwicki J.E. Non-canonical interactions between heat shock cognate protein 70 (Hsc70) and Bcl2-associated anthanogene (BAG) co-chaperones are important for client release. J. Biol. Chem. 291:19848-19857 (2016)
2. Hung W.J., Roberson R.S., Taft J., Wu D.Y. Human BAG-1 proteins bind to the cellular stress response protein GADD34 and interfere with GADD34 functions. Mol. Cell. Biol. 23:3477-3486 (2003)
Subcellular Location
Nucleus, Cytoplasm.
Synonyms
Bag 1 antibody
BAG family molecular chaperone regulator 1 antibody
BAG-1 antibody
BAG1 antibody
BAG1_HUMAN antibody
BCL 2 Associated Athanogene 1 antibody
BCL 2 associated athanogene antibody
BCL 2 binding athanogene 1 antibody
Bcl-2-associated athanogene 1 antibody
BCL2 associated athanogene 1 antibody
ExpandImages
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Western blot analysis of Bag1 on different lysates with Rabbit anti-Bag1 antibody (HA721885) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: HEK-293 cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: RAW264.7 cell lysate (20 µg/Lane)
Lane 6: Rat testis tissue lysate (40 µg/Lane)
Lane 7: Mouse testis tissue lysate (40 µg/Lane)
Predicted band size: 38 kDa
Observed band size: 35/46/52 kDa
Exposure time: 3 minutes;
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 (HA721885) 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. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Bag1 antibody (HA721885) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721885) 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 prostate cancer tissue with Rabbit anti-Bag1 antibody (HA721885) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721885) 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. -
Immunocytochemistry analysis of HeLa cells labeling Bag1 with Rabbit anti-Bag1 antibody (HA721885) 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-Bag1 antibody (HA721885) 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. -
Flow cytometric analysis of HeLa cells labeling Bag1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721885, 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"