Mcl 1 sc 12756

Unless otherwise mentioned, Western blotting was performed as described in our previous publications [21 (link)–23 (link)]. Primary antibodies for EDA2R (ab203667, 1:1000) and nephrin (ab58968, 1:1000) were purchased from Abcam (Cambridge, MA), and antibodies for Bax (sc-526, 1:1000), Bcl2 (sc-492-g, 1:1000), Mcl-1 (sc-12756, 1:1000), and actin (sc-8432, 1:3000) were from Santa Cruz Biotechnology. The antibody for cleaved caspase 3 (AF835, 1:1000) was from R&D Systems (Minneapolis, MN).

Tissue slides were dewaxed with xylene followed by rehydration with a graded alcohol series. Tissue slides were incubated in 3% (v/v) hydrogen peroxide for 10 min and then blocked with 10% BAS in PBS for 1 h. After being washed with PBS 3 times, the slides were incubated with primary antibodies (cleaved PARP 1:50, Bim 1:200 and Mcl-1 1:100) at 4°C in a humidified chamber overnight and then treated with ImmPRESS® Horse Anti-Rabbit IgG Polymer Kit (MP-7401; Vector Laboratories, Burlingame, CA) and DAB Peroxidase Substrate Kit (SK-4100; Vector Laboratories). cleaved PARP (#5625) and Bim (#2933) antibodies used in IHC were purchased from Cell Signaling Technology, Inc (Danvers, MA). Mcl-1 (sc-12756) was purchased from Santa Cruz Biotechnology (Dallas, TX).