Magic Fast™ AF647 Conjugation Kit

Creative Diagnostics provided Magic Fast™ AF647 Conjugation Kit is designed for small scale conjugation. The NHS ester groups of AF647 can react with amino group to form a stable amide group. It provides a rapid and easy process with high efficiency and fluorescent intensity to conjugate antibodies or protein to AF647. The total process completed in 3 hours and less than 30 minutes hands-on time.

Performance

• Simple and efficient labeling of antibody or protein with AF647.
• Fast and easy preparation: total process completed in 3 hours and less than 30 minutes of hands-on time.

Buffer Considerations

Procedure

(1) Equilibrate reagent to room temperature before using. Make sure all buffers are well dissolved. If not, please vortex the vial to make salts dissolved.

(2) Dissolve antibody in PBS or other buffer that do not contain amine, tris, NaN3 or glycerol. Add 10X Modifier to antibody (e.g. 1 μL of 10X Modifier for 9 μL of antibody)

(3) Spin down the vial of AF647 before using.

(4) Open the cap of the vial of AF647 and pipette antibody into the vial. Mix gently by pipetting several times until AF647 dye is well dissolved.

(5) Cover the cap on the vial and spin down the vial. Incubate in the dark at room temperature for 2 hours.

(6) Method 1: Using SpinDesalt Column to remove unconjugated AF647. Please refer to the protocol of SpinDesalt Column. Collect labeled antibody and stabilize with 1% bovine serum albumin or another stabilizer.

Method 2: Add 10X Quencher to Ab-Fluorescent mixture. Incubate in the dark at room temperature for 30 minutes. Stabilized with 1% bovine serum albumin or another stabilizer.

(7) Store the labeled protein protected from light.

(8) For protein conjugation, it can be calculated by formula below:

Quantities of protein = quantities of kit (e.g. 100 μg) x (M.W.of target protein)/(150000 (M.W.of IgG)