FIGURE 1.
Analysis of recombinant XEEL. A, Western blotting of supernatant from High FiveTM (BTI-TN-5B1-4) cells expressing the construct encoding XEEL fused to a Strep-tag II or HEK293T cells expressing the identical construct in either reducing (R) or non-reducing (oxidized (O)) conditions. A sample of hIntL-1 is included for comparison. The hIntL-1 antibody used (Proteintech, 11770-1-AP) cross-reacts with XEEL. Spent culture media from non-infected or non-transfected cells were used as negative controls. Intelectins are glycoproteins, and thus the higher molecular weight bands likely represent different glycoforms. Under non-reducing conditions, disulfide-containing proteins are nonlinear polypeptides and thus may have different mobility than their reduced, linear polypeptide counterparts. B, SDS-PAGE of thioredoxin fused to N-terminal residues 22–47 of XEEL under reducing and oxidizing conditions. The expected masses are 17, 34, and 102 kDa for monomer, dimer, and hexamer, respectively.