Caption: IP-WB analysis of K562 whole cell lysate using the MDA5 specific antibody, 700360. Lane 1 is 2.5% of five million whole cell lysate input. Lanes 2 and 3 are 50% of IP enrichment from five million whole cell lysate using normal IgG antibody and the MDA5-specific antibody, 700360. The same antibody was used to detect protein levels via Western blot. This antibody passes preliminary validation and will be further pursued for secondary validation. *NOTE* Protein sizes are taken from Genecards.org and are only estimates based on sequence. Actual protein size may differ based on protein characteristics and electrophoresis method used. Comments: Immunoprecipitation lane (3) exhibits the same double banding pattern seen in another western blot example from the antibody supplier, though they are shifted much larger in size. We believe this is due to performing the characterization using the Jess Western Blotting system, which has a documented tendency for some proteins to run differently than with more traditional methods due to their compositions. Method: immunoprecipitation Status: NotSatisfied Lab: Yeo Lab |