This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near aa 175-200 of mouse MDM2.
This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 90 kDa in size corresponding to MDM2 protein by western blotting in
Western blot using Rocklands affinity purified Anti-MDM2 (Rabbit) is shown to detect a band (arrow) corresponding to mouse MDM2 protein. Lane 1: human kidney HEK293 cells (p/n W09-000-365). Lane 2: mouse MEF cells (p/n W10-001-371). Approximately 35µg of lysate was separated by 4-20% Tris Glycine SDS-PAGE. After blocking the membrane with 5% normal goat serum, 0.5% BLOTTO (p/n B501-0500) in PBS, the membrane was probed for overnight at 4 with the primary antibody diluted to 1:500 in 1% normal goat serum, 0.1% BLOTTO in PBS. The membrane was washed and reacted with a 1:10,000 dilution of IRDye800 conjugated Gt-a-Rabbit IgG [H&L] (p/n 611-132-122) for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers indicated at the right (700 nm channel, red). IRDye800 fluorescence image was captured using the Odyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
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