Anti-Phospho-HSP70 (Tyr41) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-HSP70 (Tyr41). Anti-Phospho-HSP70 (Tyr41) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Cow)

1. Blank control (blue line): Jurkat (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-phospho-HSP70 (Tyr41) antibody (TMAB-10829), Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg /test.
2. Tissue/cell: Rat pancreas tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-phospho-HSP70 (Tyr41) polyclonal Antibody, Unconjugated (TMAB-10829) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
3. Blank control: MCF7.
Primary Antibody (green line): Rabbit Anti-phospho-HSP70 (Tyr41) antibody (TMAB-10829)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
4. Blank control: A549.
Primary Antibody (green line): Rabbit Anti-phospho-HSP70 (Tyr41) antibody (TMAB-10829)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
5. Sample: Epithelium (Mouse) Lysate at 40 μg
Primary: Anti-phospho-HSP70 (Tyr41) (TMAB-10829) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 70 kD
Observed band size: 70 kD
6. Sample:
A549 (Human) Cell Lysate at 30 μg
k562 (human) Cell Lysate at 30 μg
Primary: Anti-phospho-HSP70 (Tyr41) (TMAB-10829) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 70 kD
Observed band size: 70 kD

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human HSP70 around the phosphorylation site of Tyr41

Human

HSPA1A

Heat shock 70 kDa protein 1A

In cooperation with other chaperones, Hsp70s stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, serves as a post-attachment receptor for the virus to facilitate entry into the cell.