3 ~ 5 molecules of biotin and 1 molecule protein binding experiment

First, the experiment step:
1. Dissolve 2 to 10 mg of protein in 1 ml of BUPHTM phosphate buffer, and calculate the dissolved millimorator. If the protein contains inappropriate buffers (such as TRIS or glycine), it can be removed by dialysis or concentration in PBS. Calculate: Protein mass (Mg) / protein molecular weight (Mw) = mole number of proteins.
2. Balance the biotin to room temperature before opening. Add 2 mg sulfo-nhs-biotin in 100 ul ultrapure water, add sufficient amount of concentration of biotin, generally more than 12 times molecules for 10 mg / ml protein, more than 20 times for 2 mg / ml protein solution , Or the reagent can also be added to the protein solution in the form of a powder.
3. It is 30 minutes room temperature or 2 hours on ice.
4. Pre-wash the purified column with 30 mL PBS, the same buffer as the collection amount, and 0.5 mL or 1 ml of the separate tube is collected.
5. The protein content was determined at 280 nm absorption values.
6. Biotinylated proteins are stored at 4 ° C to use, and sodium sodium, glycerol or the like can be added.
Building Block Compounds is a term in chemistry which is used to describe a virtual molecular fragment or a real chemical compound the molecules of which possess reactive functional groups.