Search results for "Electroblotting"
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Staphylococcus aureus alpha-toxin. Production of functionally intact, site-specifically modifiable protein by introduction of cysteine at positions 6…
1993
Staphylococcal alpha-toxin, the prototype of an oligomerizing, pore-forming cytotoxin, is sensitive to biochemical modifications and cannot be labeled with biotin or fluorescein under preservation of its biological activity. In this study, we have used site-directed mutagenesis to introduce cysteine residues at positions 69, 130, and 186. Each mutant was fully and rapidly reactive with several sulfhydryl-specific reagents, indicating superficial location. Coupling of SH-groups with fluorescein-maleimide or biotin-maleimide was tolerated without loss of hemolytic activity at position 130, and the formed hexamers were visible on target cells by fluorescence microscopy and could be detected on…
Two proteases from nuclei of rat testis cells. I. Isolation
1987
Abstract Two proteases, assayed with fluorogenic peptides and tentatively designated Rc and Kc, have been isolated from nuclei of rat testis cells by differential extraction with acetic acid, removal of some proteins at pH 4.5, and polyacrylamide gel electrophoresis followed by electroblotting onto nitrocellulose paper. Protease R hydrolyzes t‐Butyl‐oxycarbonyl‐Val‐Pro‐Arg‐7‐amino‐4‐methyl‐coumarin and other peptides in which arginine is joined to 7‐amino‐4‐methyl‐coumarin by amide linkage. Protease Kc has a preference for peptides terminating in lysine‐7‐amino‐4‐methylcoumarin amide. Neither of these proteases is active against Glu‐Phe‐7‐amino‐4‐methyl‐coumarin amide or Carbobenzoxy‐Arg‐7‐…