ab53496 at 100,000 dilution staining Hsp70 + Hsc70 in mouse colon cancer tissue section by immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). A antibody amplifier™ system was used for staining. A HRP-conjugated secondary antibody was used at 1/10 dilution. 

Anti-Hsp70 antibody [BB70] (ab53496) at 1/1000 dilution + HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

Predicted band size: 70 kDa

Ab53496, at a dilution of 1/200, staining Hsp70 + Hsc70 in liver smears of rat hepatocytes by Immunohistochemistry Formalin fixed Parafin embedded tissue.

1. Control

2. Two hours after exposure tp hyperthermic stress   

ab53496 at 100,000 dilution staining Hsp70 + Hsc70 in human colon cancer tissue section by immunohistochemistry (Formalin/ PFA fixed paraffin-embedded tissue sections). A antibody amplifier™ system was used for staining.

Anti-Hsp70 antibody [BB70] (ab53496) at 1/1000 dilution + Bovine MDBK cells

Predicted band size: 70 kDa

Ab53496, at a 1/200 dilution, staining Hsp70 + Hsc70 in nuclear smears of rat hepatocytes by immunocytochemistry. 

1. Control

2.Five hours after exposure to hyperthermic stress

   

ab53496 staining Hsp70 in LLC-PK1 cells expressing Hsp70 under cold block conditions, by Immunocytochemistry/ Immunofluorescence.

Cells were fixed with 4% paraformaldehyde, permeabilized with 0.01% Triton X-100 in PBS for 4 minutes and blocked with 1% BSA/PBS for 20 minutes. Samples were incubated with primary antibody overnight at 4°C.

Overlay histogram showing Jurkat cells stained with ab53496 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab53496, 0.5µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.