Caption: IP-WB analysis of K562 whole cell lysate using the GTF2F1 specific antibody, PA5-30050. Lanes 1 and 2 are 2.5% of five million whole cell lysate input and 50% of IP enrichment, respectively, using a normal IgG antibody. Lane 3 is 50% of IP enrichment from five million whole cell lysate using the GTF2F1-specific antibody, PA5-30050. 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: We recognize the protein size discrepancy seen in the immunoprecipitation lane and believe this to be attributable to performing the characterization on the Jess Western Blotting system. This system has a documented tendency for some proteins to run differently than they would with more traditional methods due to their compositions and/or interactions with the electrophoretic components. Based on the visual and numerical data, we believe the characterizations to be real but also realize that these cases require matching knockdown-western blot characterizations and/or redoing the primary characterizations to confirm specificity. Method: immunoprecipitation Releated Sample: eCLIP:342 Status: Released Lab: Yeo Lab |