1984
DOI: 10.1073/pnas.81.8.2553
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Binding of alpha-bungarotoxin to proteolytic fragments of the alpha subunit of Torpedo acetylcholine receptor analyzed by protein transfer on positively charged membrane filters.

Abstract: Proteolytic fragments of the a subunit of the acetylcholine receptor retain the ability to bind a-bungarotoxin following resolution by polyacrylamide gel electrophoresis and immobilization on protein transfers. The a subunit of the acetylcholine receptor of Torpedo electric organ was digested with four proteases: Staphylococcus aureus V-8 protease, papain, bromelain, and proteinase K. The proteolytic fragments resolved on 15% polyacrylamide gels were electrophoretically transferred onto positively charged nylo… Show more

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Cited by 70 publications
(34 citation statements)
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“…Protein concentration was measured using the Bradford method (Bradford 1976). Denatured samples (50 µg) were subjected to 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (Laemmli 1970), and transferred to a nitrocellulose filter (Wilson et al 1984) (Bio-Rad). In the parallel experiment, we checked whether the protein samples were equally transferred to a nitrocellulose filter with staining of the filter by ponceau S solution (Sigma).…”
Section: Immunoblot Analysismentioning
confidence: 99%
“…Protein concentration was measured using the Bradford method (Bradford 1976). Denatured samples (50 µg) were subjected to 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (Laemmli 1970), and transferred to a nitrocellulose filter (Wilson et al 1984) (Bio-Rad). In the parallel experiment, we checked whether the protein samples were equally transferred to a nitrocellulose filter with staining of the filter by ponceau S solution (Sigma).…”
Section: Immunoblot Analysismentioning
confidence: 99%
“…The protein concentration was determined using the Bradford method. Twenty-five micrograms denatured samples were subjected to 10% SDS-PAGE (Laemmli 1970), and transferred to a nitrocellulose filter (Wilson et al 1984) (Bio-Rad). The filter was incubated with a rabbit anti-GLUT1 antiserum ) at a dilution of 1:8000, followed by a second incubation with goat antirabbit immunoglobulin G conjugated to horseradish perioxidase (Bio-Rad) at a dilution of 1:16 000.…”
Section: Immunoblot Analysismentioning
confidence: 99%
“…The analysis of a-bungarotoxin (a-BTX) binding to proteolyzed or deglycosylated AcChoR has also been employed in search of the AcCho-binding site (8)(9)(10)(11)(12). Protein blotting has been found to be particularly useful in this respect.…”
mentioning
confidence: 99%
“…We have synthesized two segments of the Torpedo a subunit, peptides [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] Merrifield (18). Protected amino acids were either purchased from Peptides Japan (Osaka) or generously provided by I. Jacobson (Department of Biophysics, The Weizmann Institute of Science, Rehovot).…”
mentioning
confidence: 99%