Bcl xl antibody

Total cell lysates of paclitaxel-treated or control LNCaP or PC3 cells were prepared in Chaps buffer (5mM MgCl₂, 137mM NaCl, 1mM EDTA, 1mM EGTA,1% Chaps, 20mM Tris–HCl (pH 7.5)), and protease inhibitors. About 500 μg of proteins were immunoprecipitated with Bcl-xl antibody (Abcam) at 4°C for 2 h. Immunoprecipitates were captured by 50 μl of the slurry of protein A-magnetic beads (Millipore) in Chaps buffer at 4°C for overnight. Immunoprecipitates were then recovered by magnetic stand and washed three times in 1% Chaps buffer. Immunoprecipitates were eluted by SDS-PAGE sample buffer, were subjected to SDS-PAGE and then immunoblotting.

Bcl-xL antibody (1:1000, Cat#: ab32370) and VASH1 (1:1000, Cat#: ab199732) were purchased from Abcam. Bcl-2 antibody (1:1000, Cat #: 15071S), and Cleaved Caspase-3 (1:1000, Cat#: 9661S) were purchased from Cell Signaling Technologies. α-tubulin (1:1000, Cat#: T6199), FLAG (1:1000, Cat#: F3165), Tyr-Tub antibody (1:1000, Cat# MAB1864-I), and VASH2 (1:1000, Cat#: MABC536) were purchased from Sigma. GAPDH (1:5000, Cat#: sc-32233) was purchased from Santa Cruz. SVBP (CCDC23, 1:1000, Cat#: PA5-52569) was purchased from Invitrogen. The deTyr-Tub antibody was developed by Takashi Hotta and Ryoma Ohi [17 (link)]. This antibody is now commercially available through RevMab under catalogue number RM444 (1:10000). Alexa Fluor 568 (1:1000, Cat#: A11011), Alexa Fluor 594 (1:1000, Cat#: A11012) and Hoechst 33258 (1:5000, Cat#: H3569) were purchased from Invitrogen (Waltham, MA, USA).