T1 and T2 cells were lysed and protein was isolated using Radioimmunoprecipitation assay (RIPA) lysis buffer (R0278-50ML, Sigma-Aldrich) with protease inhibitor (P8340-5ML, Sigma-Aldrich). The protein concentrations were quantified using BCA assay (23225, Thermo Fisher Scientific). 25 µg of total protein lysate was loaded on to 10% SDS gel and western blotting was performed as previously described (42 (link)). Antibodies used in the study are listed in Supplementary Table 5.
Thankamony A.P., Ramkomuth S., Ramesh S.T., Murali R., Chakraborty P., Karthikeyan N., Varghese B.A., Jaikumar V.S., Jolly M.K., Swarbrick A, & Nair R. (2023). Phenotypic heterogeneity drives differential disease outcome in a mouse model of triple negative breast cancer. Frontiers in Oncology, 13, 1230647.
Corresponding Organization : Centre For Human Genetics
Other organizations :
Manipal Academy of Higher Education, Garvan Institute of Medical Research, The Kinghorn Cancer Centre, Indian Institute of Science Bangalore, UNSW Sydney, St Vincent's Clinic
Protein expression levels measured by Western blotting
control variables
RIPA lysis buffer with protease inhibitor used for protein extraction
Protein concentration quantified using BCA assay
25 µg of total protein lysate loaded onto 10% SDS gel
Western blotting protocol as previously described
controls
Positive and negative controls not explicitly mentioned
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