Using C2C12 myotubes, I plan to carry out the glucose absorption assay (2-NBDG method). But, I am not understanding the procedure which they followed. A KRPH buffer formula consisting of 20 mM HEPES, 5 mM KH2PO4, 1 mM MgSO4, 1 mM CaCl2, 136 mM NaCl, and 4.7 mM KCl at pH 7.4 is used in certain publications, but pyruvate is used in publications.

1. What is the ideal KRPH buffer composition?

2. Is it required to lyse the cells in order to use a microplate reader to measure the absorbance after adding 2-NBDG? (if 'yes' means, the reason)

Could someone help me to solve it?

Thanks in advance....