Anti-Hsc70 antibody [1B5] (ab19136)

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ab19136 staining Hsc70 in Human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/150) for 12 hours at 4°C. A Biotin-conjugated Goat anti-rat polyclonal (1/200) was used as the secondary antibody.

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Blocking: 5% milk for 1 hour at 22°C.

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ab19136 at 1/150 dilution staining human HeLa cells by ICC/IF. The cells were heat shocked (2 hours at 42.5°C) and paraformaldehyde fixed prior to incubation with the antibody for 1 hour at 37°C. An Alexa-Fluor ^® 568 conjugated goat anti-rat antibody was used as the secondary. The cells were also stained with DAPI to visualize the nuclei.

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Overlay histogram showing HeLa cells stained with ab19136  (red line). The cells were fixed with 80% methanol (5 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 (ab19136, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.