Sulfo-NHS-LC-LC-Biotin

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
Calculation: To achieve the best results, use a biotin concentration of ≥ 12 times molar excess for the 10 mg/mL protein solution, or ≥ 20 times molar excess for the 2 mg/mL protein solution.
Calculate the number of millimoles of biotin reagent to be added to the reaction to achieve a 20-fold molar excess:
mL of protein × (mg of protein/mL of protein) × (mmol of protein/mg of protein) × (20 mmol of protein/mmol of protein) = mmoL of biotin.
20 = The recommended molar excess of biotin for a 2 mg/ml protein sample.
2. The volume (in microliters) of the 10 mM biotin solution to be added to the reaction:
mmoL biotin × (1000000 μL/L) × (L/10 mmoL) = μL biotin Preparation of Biocytin-Linked Antibody 1 (bio-Ab1)^[1] Add 200 μL of the antibody (0.83 μM, dissolved in PBS) and 1.66 μL of 2 mM Sulfo-NHS-LC-LC-Biotin (dissolved in DMSO) to a reaction vessel, and let it react at room temperature for 1 hour with continuous shaking.
2. The reaction mixture was subjected to buffer exchange and then purified using a Zeba rotary desalination column (with a molecular weight cut-off of 40K, balanced with PBS) to remove the unbound biotin.
3. Stabilize the purified biotinylated antibody (bio-Ab1) in PBS buffer containing 0.1% (w/v) BSA, and store it at 4℃.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.