Western blot using PPP1R13L polyclonal antibody (Cat # PAB10243) shows detection of a band at ~100 KDa (arrowhead) corresponding to isoform 1 of PPP1R13L in MCF-7 whole cell lysates. Preincubation with immunizing peptide blocks specific band staining (data not shown). Approximately 35 ug of lysate was separated by 4-20% Tris Glycine SDS-PAGE. After blocking, the membrane was probed with the primary antibody diluted to 1:1,500 in 5% BLOTTO/PBS overnight at 4°C. The membrane was washed and reacted with a 1:10,000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers. IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.
Immunohistochemistry of PPP1R13L polyclonal antibody (Cat # PAB10243) shows strong cytoplasmic and membranous staining of myocytes in human heart tissue. Tissue was formalin-fixed and paraffin embedded. Brown color indicates presence of protein, blue color shows cell nuclei. Personal Communication, Kenneth Wester, www.proteinatlas.org, Uppsala, Sweden.