Studies of phosphorylation in rat mast cell. (Supplement-1). Demonstration of triphosphoinositide(TPI) in rat mast cell granule.:—<I>demonstration of triphosphoinositide (TPI) in rat mast cell granule</I>—
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Granules were isolated from sonicated rat mast cells on Percoll gradient and incubated with [γ<SUP>32</SUP>P] ATP in the presence of Mg<SUP>2+</SUP> and exogenous diphosphoinositide. After stopping the reaction, lipid extraction of the intact membrane granules was performed with acidic medium and phospholipids were separated by thin layer chromatography on 1% (w/v) oxalic acid and potassium oxalate impregnated silica gel plates. Considerable amounts of radioactivity were found to be incorporated into TPI on both plates. Extensive washing of the granules did not significantly affect the <SUP>32</SUP>P incorporation into TPI. The radioactivity in TPI was associated with the granule themselves and it is apparent that TPI is produced in rat mast cell granule.
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