Active Chondroitin Sulfate Proteoglycan 4 (CSPG4)

Figure. The binding activity of MCSP with GAL8.
Melanoma-associated chondroitin sulfate proteoglycan (MCSP) is a chondroitin sulfate proteoglycan in humans. CSPG4 plays a role in stabilizing cell-substratum interactions during early events of melanoma cell spreading on endothelial basement membranes. It represents an integral membrane chondroitin sulfate proteoglycan expressed by human malignant melanoma cells. Besides, Galectin-8 (GAL8) has been identified as an interactor of MCSP, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MCSP and recombinant human GAL8. Briefly, MCSP were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to GAL8-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MCSP pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of MCSP and GAL8 was shown in Figure 1, and this effect was in a dose dependent manner.

Figure. Western Blot