A simplified solid-phase assay for the quantitation of native membrane proteins. Application to the measurement of EGF receptor induction by dexamethasone.
Direct application of membrane fractions to a nitrocellulose support without previous solubilization, denaturation or fixation of the antigen permits the quantitation of antibodies binding to denaturation sensitive epitopes. In experiments reported here, this method was used to determine the rate of EGF receptor expression on squamous carcinoma cell lines. The assay revealed that dexamethasone treatment leads to an eight-fold increase of EGF receptor protein expression on C4-I cervical carcinoma cells. The data, when compared to the results obtained in immunoprecipitation experiments, suggest that this simple protocol yields reliable and precise quantitative data. The simplicity of the method permits simultaneous testing of large sample numbers with various antibodies.
Von Knebel Doeberitz
M
M
Institut für Virusforschung/ATV, Deutsches Krebsforschungszentrum, Heidelberg, F.R.G.
Drzonek
H
H
Koch
S
S
Becker
C M
CM
D002294:Carcinoma, Squamous Cell / Q000032:analysis; D003907:Dexamethasone / Q000494:pharmacology*; D066246:ErbB Receptors / Q000032:analysis*; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D014407:Tumor Cells, Cultured
D003907:Dexamethasone; D066246:ErbB Receptors