Figure 3.

Purification and detection of CD45 and its isoforms as Jacalin receptors on Raji B cells. (a, b) Purification and detection of Jacalin receptors by affinity chromatography, and Western and lectin blotting. Jacalin receptors were purified from human Raji B-cell membrane proteins on a Jacalin-agarose affinity column and fractionated on a 5–20% reducing gradient sodium dodecyl sulphate–polyacrylamide gel electrophoresis gel, and then transferred to nitrocellulose membranes, followed by pan-CD45 Western blot (a) and Jacalin lectin blot (b) detection. The arrows indicate the purified Jacalin receptors on Raji B cells. The molecular weight markers are shown on the left. (c) Expression of CD45 isoforms on Raji B cells revealed on flow cytometry analysis. Raji B cells were stained with fluorescein isothiocyanate-conjugated anti-human CD45RA, CD45RB and CD45RO monoclonal antibodies, respectively, and then analysed by flow cytometry. As a negative control, the autofluorescence of the cells was measured (grey area).