bsm-52241R
[Primary Antibody]
Hsp70 (2G2) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Hsp70
Clonality: Monoclonal
Isotype: IgG
Swiss Prot: P17879, Q61696
Source: Recombinant Mouse Hsp70 from 400 amino acid to C-terminus
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Background:
Molecular chaperone implicated in a wide variety of cellular processes, including protection of the proteome from stress, folding and transport of newly synthesized polypeptides, activation of proteolysis of misfolded proteins and the formation and dissociation of protein complexes. Plays a pivotal role in the protein quality control system, ensuring the correct folding of proteins, the re-folding of misfolded proteins and controlling the targeting of proteins for subsequent degradation. This is achieved through cycles of ATP binding, ATP hydrolysis and ADP release, mediated by co-chaperones. The co-chaperones have been shown to not only regulate different steps of the ATPase cycle, but they also have an individual specificity such that one co-chaperone may promote folding of a substrate while another may promote degradation. The affinity for polypeptides is regulated by its nucleotide bound state. In the ATP-bound form, it has a low affinity for substrate proteins. However, upon hydrolysis of the ATP to ADP, it undergoes a conformational change that increases its affinity for substrate proteins. It goes through repeated cycles of ATP hydrolysis and nucleotide exchange, which permits cycles of substrate binding and release. The co-chaperones are of three types: J-domain co-chaperones such as HSP40s (stimulate ATPase hydrolysis by HSP70), the nucleotide exchange factors (NEF) such as BAG1/2/3 (facilitate conversion of HSP70 from the ADP-bound to the ATP-bound state thereby promoting substrate release), and the TPR domain chaperones such as HOPX and STUB1 (PubMed:24012426, PubMed:26865365, PubMed:24318877). Maintains protein homeostasis during cellular stress through two opposing mechanisms: protein refolding and degradation. Its acetylation/deacetylation state determines whether it functions in protein refolding or protein degradation by controlling the competitive binding of co-chaperones HOPX and STUB1. During the early stress response, the acetylated form binds to HOPX which assists in chaperone-mediated protein refolding, thereafter, it is deacetylated and binds to ubiquitin ligase STUB1 that promotes ubiquitin-mediated protein degradation (PubMed:27708256). Regulates centrosome integrity during mitosis, and is required for the maintenance of a functional mitotic centrosome that supports the assembly of a bipolar mitotic spindle (PubMed:27137183). Enhances STUB1-mediated SMAD3 ubiquitination and degradation and facilitates STUB1-mediated inhibition of TGF-beta signaling (PubMed:24613385). Essential for STUB1-mediated ubiquitination and degradation of FOXP3 in regulatory T-cells (Treg) during inflammation (PubMed:23973223). Negatively regulates heat shock-induced HSF1 transcriptional activity during the attenuation and recovery phase period of the heat shock response (PubMed:9499401).
Size: 100ul
Concentration: 1ug/ul
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Lane 1: MCF-7; Lane 2: HCT116; Probed with Hsp70 (2G2) Monoclonal Antibody (bsm-52241R) at 1:1000 overnight at 4°C followed by a conjugated secondary antibody for 60 minutes at 37°C.
Flow cytometric analysis of Hela cells with Hsp70 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.
IF(ICC) staining with Hsp70 (2G2) Monoclonal Antibody (bsm-52241R) at 1:100 in HeLa cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
Paraformaldehyde-fixed and paraffin-embedded Human breast carcinoma tissue incubated with Hsp70 (2G2) Monoclonal Antibody (bsm-52241R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed and paraffin-embedded Mouse prostate tissue incubated with Hsp70 (2G2) Monoclonal Antibody (bsm-52241R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed and paraffin-embedded Mouse testis tissue incubated with Hsp70 (2G2) Monoclonal Antibody (bsm-52241R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Hsp70 (2G2) Monoclonal Antibody, Unconjugated (bsm-52241R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Hsp70 (2G2) Monoclonal Antibody, Unconjugated (bsm-52241R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Mouse Testis lysates; Lane 2: Mouse NIH/3T3 cell lysates; Lane 3: Rat Liver lysates; Lane 4: Rat Testis lysates; Lane 5: Human Hela cell lysates; Lane 6: Human Jurkat cell lysates probed with Hsp70 Monoclonal Antibody, Unconjugated (bsm-52241R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Jurkat cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Hsp70 (2G2) Monoclonal Antibody(bsm-52241R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (HSP70) monoclonal Antibody, Unconjugated (bsm-52241R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.