Appendix 6 of Sharma, Mol Vis 2018; 24:801-817.
Human serum proteins (15 μg each), human red blood cell (RBC) lysate or HEK293A cell lysate (50 μg), and SRA 01/04 cell (50 μg each) and human whole lens (50 μg each) lysates were analyzed by SDS–PAGE and western blotting performed with the antibody specified at the top of each panel. The ~26 kDa band detected in serum with the anti-APOA1 antibody corresponds to the expected size of secreted APOA1 and the faint ~50 kDa band is likely a dimer. The faint bands seen in SRA 01/04 cells are most likely non-specific. The ~41 kDa band detected with the anti-APOA4 antibody corresponds to the expected size of secreted APOA4, the bands between 60 and 75 kDa in SRA 01/04 cells are likely non-specific. The ~20 kDa band detected with the anti-CRYAA antibody in human lens corresponds to the expected size of CRYAA monomer; the higher molecular mass bands likely represent the expected oligomeric forms of the protein. The ~55 kDa protein band in human serum detected with the anti-FGB antibody corresponds to the expected size of FGB. The <15 kDa protein band detected with the anti-HB antibody in RBCs likely represents the monomeric HB subunits. The monomeric subunits migrated slightly faster than their expected sizes of 15 to 17 kDa. The higher ~30 and ~50 kDa protein bands detected in the RBC lysate likely respectively represent di- and tetra- meric forms of HB. The faint 75–100 kDa bands in SRA 01/04 cells are likely non-specific. The ~25 kDa band in SRA 01/04 cells detected with the anti-HSPB1 antibody likely represents HSPB1 but the protein migrated slightly faster than its expected size of 27 kDa. The <25 kDa band in HEK293A and SRA 01/04 cells detected with the anti-PRDX2 antibody likely represents PRDX2 monomer, however, migration of the protein was slightly retarded than its expected size of 22 kDa. The <50 kDa bands in HEK293A and SRA 01/04 cells likely represent PRDX2 dimers. The faint ≥75 kDa bands in the two cell types are most likely none specific. None of the proteins except CRYAA was detected in human lens. Numbers along the panels indicate molecular masses of protein standards in kilo Daltons. Serum, human serum; SRA, SRA 01/04 cells; blood, red blood cells; 293A, HEK293A cells. To access the data, click or select the words “Appendix 6.”