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Our Abpromise guarantee covers the use of ab19136 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 73 kDa.|
|IP||Use a concentration of 5 µg/ml.|
This image is courtesy of an anonymous Abreview
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.
This image is courtesy of an anonymous abreview.
Blocking: 5% milk for 1 hour at 22°C.
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.
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.
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