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Our Abpromise guarantee covers the use of ab9063 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent dilution.|
|ICC/IF||Use a concentration of 5 µg/ml.|
14-3-3 is required for CT-Raf kinase activity. 14-3-3 can be removed from CT-Raf in vitro with the detergent Empigen-BB. Sf9 cells were infected with recombinant baculoviruses encoding either GST or a GST–CT-Raf fusion protein. Forty-eight hours after infection, the GST or GST–CT-Raf proteins were isolated from NP-40 lysates by using glutathione-agarose beads.
The bead-bound complexes were then washed with either NP-40 lysis buffer (lanes 1 and 2) or NP-40 lysis buffer containing 1% Empigen-BB (lane 3), resolved by SDS-PAGE, transferred to nitrocellulose, and developed with an antibody to either 14-3-3 (upper panel) or Raf (lower panel).
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