Cyclin E1 (HE12) Mouse mAb #4129

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	-Mouse (1:1000)

Protocol tips
-Mouse (1:1000)

Publication protocol

The following antibodies were used for Western blotting: APC11 (1/500, ab57158) and MYC (1/5,000, ab32072), Abcam; APC3 (1/1,000, 610454), cyclin A (1/1,000, 611269), Mcl‐1 (1/500, 559027) and Nek2A (1/500, 610593), BD Transduction Laboratories; APC8 (1/1,000, A301‐181A) and Fbw7 (A301‐720A‐1), Bethyl Laboratories; APC2 (1/1,000, 12301S) and cyclin E (1/1,000, 4129S), Cell Signaling Technologies; MPM2 (1/1,000, 05‐368), Millipore; APC10 (1/1,000, sc‐166790), Cdc20 (1/1,000, sc‐13162) and cyclin B1 (1/2,000, sc‐752), Santa Cruz Biotechnology; Actin (1/4,000, A2066) and Mad2 (1/1,000, PA5‐21594), Sigma Aldrich; Cdh1 (1/500, MS1116‐P0), Thermofisher. Antibodies were used according to standard protocols except for Fbw7 where blocking and primary antibody incubation were carried out in ReliaBLOT (Bethyl Laboratories). Histone H3 phospho‐Ser10 (05‐806), Millipore, was used for flow cytometry as described previously (Harley et al, 2010). Fbw7 (A301‐721A‐1), Bethyl Laboratories, was used for immunoprecipitation. Doxycycline, thymidine, paclitaxel and reversine were purchased from Sigma Aldrich. Nocodazole (Calbiochem) was routinely used at 830 nM apart from assaying cyclin B1 degradation where it was used at 33 nM. ProTAME was initially provided by Dr Randall King (Dept. of Cell Biology, Harvard University) and subsequently purchased from Boston Biochem.

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Manufacturer protocol

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