Labeling Procedure for Reduced IgG1. Prepare 10 mM of the biotin labeling reagent using DMSO.2. Prepare 100 μl of 1 mg per ml reduced IgG/ml buffer solution that does not contain any large molecules with SH groups. Reduced IgG can be prepared by TCEP (tricarboxyethylphosphine), DTT, or 2-mercaptoethylamine.3. Add 1-5 μl of biotin labeling reagent DMSO solution to the IgG buffer solution and incubate at 37ºC for 1 hour.4. Remove excess biotin labeling reagent using a gel column or a Filtration tube.5. Prepare solutions for further experiments using an appropriate buffer, such as PBST (0.05% Tween 20/PBS).
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