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文献和实验Coupling Peptides To Resin For Affinity Purification
5% ethylene diamine in water. Incubate 15' at RT. Wash with 10 volumes water. At this point you have amino-affigel. Wash with 3 volumes of 0.1 M NaPi pH 7.8. Resuspend resin in 0.1 M NaPi pH 7.8. Add iodoacetic acid-NHS ester, and incubate
Coupling Peptides To Resin For Affinity Purification
volumes water. At this point you have amino-affigel. Wash with 3 volumes of 0.1 M NaPi pH 7.8. Resuspend resin in 0.1 M NaPi pH 7.8. Add iodoacetic acid-NHS ester, and incubate for 10 min at RT. The resin has about 10 5mol groups/ml, so add
Revised amino-allyl labeling protocol
, all Tris must be removed from the reaction to prevent the monofunctional NHS-ester Cy-dyes from coupling to free amine groups in solution. 3. Add 450 m L water to each reaction. C. Cleanup Add 500 m L neutralized, diluted reaction
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