Assayed samples using gamma and scintillation counters.
The major proteins responsible for potassium uptake in these cells have been established to be the NaKCl cotransport plasma membrane molecule, inhibited by bumetanide, and the NaK-ATPase pump protein, inhibitable by ouabain.
Since the majority of uptake is abolished when both bumetanide and ouabain are added, ouabain-insensitive counts were usually used to asses NaKCl cotransport function.
Showed b
-AR
and dopamine receptor
regulation of NaKCl cotransport function
via PKA and cAMP using Radioimmunoassay.
Determined NaKCl cotransport protein levels using 7.5% SDS-PAGE electrophoresis and western blot.
Showed importance of protein kinase C (PKC) using phorbol, 12-myristate, 13-acetate (PMA).
In the CPE cultured cells, protein phosphatase 1 (PP1) inhibition by calyculin A can induce catastrophic whole cell translocations at levels above 50nM, but starting from 1nM, where cotransport activity is basal, it increases by 60% at 10nM activity.
Of course, at calyculin levels >50nM, cotransport decreased, but this was probably due to reduced protein levels, as shown by a modified Lowry assay in the method of Peterson (Analytical Biochemistry 83, 346(1977)), via the loss of cells, as observed in light micrographs produced by a Nikon microphotographic system. wrote quiz and test; gave quarter grades. Numerous short term MS assignments.
Early Field Experience at Mission HS: 9 th grade general science and 12 th grade Health class.
Susan Boshoven, ESL/Science Teacher.
Maintained contact information database.
utilizing LAN. Operated Multimek automated pipetter and Carl Creative plate stacker robots that required VB interface.
Monitored increased pipette tip use due to throughput increase.
Interpreted Bylaws on the central administrative committee. Central food warehouse. House committee minutes secretary for 2 years.
General office and maintenance work.