Characterization of temperature-dependent p53 protein conformation in RPMI 8226 and HS Sultan MM cell lines.

RPMI 8226 and HS Sultan cells were cultured at 37°C (A, C) or 30°C (B, D). Total tumor cell lysates (1 mg/sample) were immunoprecipitated (10 μg) with either Ab-5 anti-wtp53 (Pab 1620) mAb or Ab-3 anti-mtp53 (Pab 240) mAb and immunoblotted (10 μg) with DO-1 HRP-conjugated anti-ptp53 mAb (A, B). Tumor cells were also mock- or γ-irradiated (2 cGy) and cultured at 37°C (C) or at 30°C (D) for 2 hours to induce wtp53 expression. Total cell lysates (30 μg/sample) of mock- or γ-irradiated tumor cells were resolved by SDS-PAGE and immunoblotted (10 μg) with DO-1 HRP-conjugated anti-ptp53 mAb or with Ab-1 anti-actin mAb.