Hrp conjugated anti rabbit 7074

The procedures used have been described (Mann & Adegoke, 2021 (link)). Briefly, tissue samples were homogenized in 7X complete buffer: 20 mM HEPES, 2 mM EGTA, 50 mM NaF, 100 mM KCI, 0.2 mM EDTA, and 50 mM B‐Glycerophosphate, supplemented just before use with 10 μL/mL of each of protease inhibitor (Sigma Aldrich, #P8340) and phosphatase inhibitor cocktails (Sigma Aldrich, #P5726), 0.2 M sodium vanadate (2.5 μL/mL), 1 M DTT (1 μL/mL) and 0.2 M benzamidine (5 μL/mL). Homogenates were then centrifuged at 1000 g for 3 min at 4°C. The resulting supernatant was removed and centrifuged again at 10,000 g for 30 min at 4°C. Protein concentrations in the supernatant were determined using the Pierce BCA Protein Assay Kit (Thermo Scientific, #23225, Waltham, MA). Equal amounts of protein (~25 μg) were separated on 10% or 15% SDS‐PAGE gels and transferred onto polyvinylidene difluoride (PVDF) membranes (0.2 μM, BIO‐RAD). Incubation of membranes in primary (Table S1) and secondary antibodies (HRP‐conjugated anti‐rabbit (#7074) or anti‐mouse (#7076), Cell Signaling Technology, Danvers, MA), imaging and quantification of data were as described (Jeganathan et al., 2014 (link); Mora & Adegoke, 2021 (link); Zargar et al., 2011 (link)).