The whole-cell patch-clamp technique has been applied to elucidate the effects of H2O2 and the SH-group reagent 2,2'-dithio-bis(5-nitropyridine) (DTBNP) on Ca2+ currents of mouse pancreatic B-cells. We also tested the effect of DTBNP on the whole-cell K+ ATP current. DTBNP (20 and 100 mumol/l) blocked both the current through Ca2+ channels (with Ba2+ as the charge carrier) and the K+ATP current in a concentration-dependent manner. The results suggest that the channels themselves or regulating proteins possess free SH-groups which may be of functional relevance. However, H2O2 did not influence the Ca2+ channel currents under standard whole-cell conditions as well as when recorded with the perforated patch technique. These results indicate that the two agents act differently on ion channel currents in B-cells although both are known to oxidize SH-groups.