Detection of electrophoretic variants of Notch, PS integrin, and DROP-1 proteins in Drosophila following extraction in guanidine hydrochloride.
A method is presented for the rapid extraction of proteins from Drosophila tissues. This method involves lysis of embryos in high concentrations of guanidine hydrochloride, followed by ultracentrifugation in a guanidine hydrochloride step gradient. Several membrane-associated antigens, including Notch and the beta subunit of PS integrin are enriched in this preparation. The quantity of the proteoglycan, DROP-1, obtained from Drosophila eggs and testes was also greatly improved by the guanidine hydrochloride extraction method. This method should prove useful in the isolation and characterization of many Drosophila antigens, particularly those associated with cell membranes.
Zackson
S L
SL
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Graner
M W
MW
Karr
T L
TL
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000196:embryology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004625:Embryo, Nonmammalian / Q000737:chemistry; D014644:Genetic Variation; D019791:Guanidine; D006146:Guanidines; D007301:Insect Hormones / Q000235:genetics / Q000302:isolation & purification*; D039001:Integrin alpha Chains; D016023:Integrins / Q000235:genetics / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification*; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000235:genetics / Q000302:isolation & purification*; D051880:Receptors, Notch
D029721:Drosophila Proteins; D006146:Guanidines; D007301:Insect Hormones; D039001:Integrin alpha Chains; D016023:Integrins; D008565:Membrane Proteins; C497759:N protein, Drosophila; C082187:P-1 protein, Drosophila; D011509:Proteoglycans; D051880:Receptors, Notch; C082188:if protein, Drosophila; D019791:Guanidine