Rat anti bim 3c5

FO B cells (B220⁺CD21^medCD23⁺AA4^–) and pB1a (CD19⁺B220^loCD5⁺) from 2-mo-old C.B17 mice and CLL/lymphoma (CD19⁺B220^loCD5^+/lo in TC+ WT or B220^loCD5^+/lo ATAid⁺ in TC^– or TC⁺ ATAμκTg mice) were purified by cell sorting (2 × 10⁶ cells/tube), and cell lysates were subjected to SDS-PAGE. Anti–c-Myc, anti–Mcl-1, and HRP goat anti–rabbit IgG antibodies were all obtained from Cell Signaling Technologies. Anti–Bcl-2 (BioLegend) with HRP rat anti–mouse IgG1 (Southern Biotech) was also used. Rat anti-Bim 3C5 and rat anti-Bmf 17A9 were both purchased from Enzo Life Sciences, together with HRP-goat anti–rat (Cell Signaling Technology). Rabbit anti-Bmf (Abcam; ab9655) was used together with HRP goat anti–rabbit IgG (Cell Signaling Technology). Anti-b actin was obtained from Bethyl Labs.

Details on sample preparation and SDS-PAGE can be found in the Online Supplementary Material. Immunoprobing was performed with the following antibodies: mouse anti-tubulin (B-5-1-2; Sigma-Aldrich); rabbit anti-p21 (C-19; sc-397) and rabbit anti-CHOP (B-3) (Santa Cruz Biotechnologies); rabbit anti-cleaved caspase-3(D175) (5A1E) and rabbit anti-PUMA (#7467) from Cell Signaling Technology; rat anti-BIM (3C5, Enzo Life Sciences); and rabbit anti-p19 INK4d (ab102842, Abcam). Rabbit monoclonal anti-BOK, RabMab BOK-1-5 [15 (link)]. IRDye^® 800CW and 600CW near-infrared fluorochrome-conjugated secondary antibodies were purchased from LI-COR Biotechnology (Bad Homburg, DE). Quantitative western blot analysis was performed using an Odyssey Fc digital imaging system from LI-COR Biotechnology. Alternatively, horseradish peroxidase-coupled secondary antibodies (Jackson ImmunoResearch Europe Ltd.) were used and signals detected on an Odyssey Fc system by enhanced chemiluminescence (Luminata Forte, Merck Millipore). Data was quantified using Image Studio 3.1.4 software (LI-COR).