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- W1034885943 abstract "This chapter highlights the usage of avidin–iminobiotin complexes for purifying plasma membrane proteins. The basis of the technique is the covalent attachment of compounds containing 2-iminobiotin, the cyclic guanidino analog of biotin, onto cell surface proteins. The tagged species are then isolated by virtue of the unique interaction between the covalently attached ligand and its binding protein, avidin. The pH-dependent interaction of 2-iminobiotin with avidin makes the recovery possible. The specific site of covalent attachment can be varied depending on the congener of 2-iminobiotin used and the choice of prior chemical or enzymatic treatment of the target membranes. The method permits labeling under native conditions in which cell membrane integrity is maintained. Prior to retrieval of 2-iminobiotin-derivatized proteins, cells are subjected to detergent solubilization. The detergent type and concentration must be determined for the particular cell system under study. 2-iminobiotin tagged cells are solubilized using 500 μl of solubilization buffer for 100 μl packed washed cells. Solubilization is allowed to proceed for 30 min at 4° with occasional vortexing." @default.
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- W1034885943 date "1986-01-01" @default.
- W1034885943 modified "2023-10-18" @default.
- W1034885943 title "[15] Use of avidin-iminobiotin complexes for purifying plasma membrane proteins" @default.
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- W1034885943 doi "https://doi.org/10.1016/0076-6879(86)22153-9" @default.
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