Human IRS1 was derived in Human HEK293T cell line. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix (Sigma), 1mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate.
This gene encodes a protein which is phosphorylated by insulin receptor tyrosine kinase. Mutations in this gene are associated with type II diabetes and susceptibility to insulin resistance.
The cell lysate is provided in RIPA buffer.
Upon receiving, store the sample at -20 centigrade. Lysate samples are stable for 12 months from date of receipt when stored at -20 centigrade. Avoid repeated freeze-thaw cycles.Lysate samples can be diluted with 2xSDS Sample Buffer provided. After dilution, the protein sample should be aliquoted and stored at -20 centigrade for long term storage. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.