Anti-HSP70 Polyclonal Antibody is a Rabbit antibody targeting HSP70. Anti-HSP70 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-HSP-70 Polyclonal Antibody, Unconjugated (TMAB-00893) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control (blue line): Jurkat (blue).
Primary Antibody (green line): Rabbit Anti-HSP70 antibody (TMAB-00893)
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.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (HSP70) polyclonal Antibody, Unconjugated (TMAB-00893) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
4. Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (HSP70) polyclonal Antibody, Unconjugated (TMAB-00893) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
5. Sample:
Lane 1: Testis (Mouse) Lysate at 40 μg
Lane 2: Epididymis (Mouse) Lysate at 40 μg
Lane 3: Seminal vesicle (Mouse) Lysate at 40 μg
Lane 4: Liver (Mouse) Lysate at 40 μg
Lane 5: NIH/3T3 (Mouse) Cell Lysate at 30 μg
Lane 6: Testis (Rat) Lysate at 40 μg
Lane 7: Spleen (Rat) Lysate at 40 μg
Lane 8: Hela (Human) Cell Lysate at 30 μg
Lane 9: Jurkat (Human) Cell Lysate at 30 μg
Lane 10: HepG2 (Human) Cell Lysate at 30 μg
Lane 11: A549 (Human) Cell Lysate at 30 μg
Lane 12: MCF-7 (Human) Cell Lysate at 30 μg
Primary:
Anti-HSP70 (TMAB-00893) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 70 kDa
Observed band size: 68 kDa
6. 25 μg total protein per Lane of various lysates probed with HSP70 polyclonal antibody, unconjugated (TMAB-00893) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

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

Polyclonal

KLH conjugated synthetic peptide: human HSP70

Human

Heat Shock Proteins,Tumor biomarkers