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Our Abpromise guarantee covers the use of ab47455 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Electron Microscopy||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 70 kDa.|
|IP||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
All are cancer cell lines.
Western blot of Hela cell heat shock lysate, using HRP conjugated goat anti-mouse secondary antibody for detection. (1µg/mL ab47455 was used to detect in 10µg of heat shocked HeLa cell lysate)
ab47455 staining Hsp70 in mouse colon cancer tissue section by Immunohistochemistry (Bouin's fixed paraffin embedded tissue sections). The primary antibody was diluted at 1/100,000. A Fluorophore conjugated goat anti mouse was used as secondary. An antibody amplifier™ system was used for staining.
Overlay histogram showing HeLa cells stained with ab47455 (red line). The cells were fixed with 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 (ab47455, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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