单克隆抗体胶乳包被方案

Reagent and equipment requirement

Binding Buffer: 0.1 M MES buffer pH 5.0 Washing Buffer: TBS-T (25 mM Tris-HCl, pH 7.2, 0.15 M NaCl, 0.05 % Tween20)

Coupling Reagent: 10 mg/mL EDC in ice-cooled Binding Buffer, prepared just before the coupling reaction Equipment: Magnetic separator. Vortex tube mixer. Tube rotator.

1. Suspend the Magnosphere™ MS300/Carboxyl beads well using Vortex mixer and put 1 mL of the suspension (i.e., 10 mg beads) into a microtube.

2. Place the tube on a magnetic separator for 1 minute (or longer if needed) and remove the supernatant carefully.

3. Add 1 mL of Binding Buffer and suspend the beads by vortexing. Then, remove the supernatant as in step 2.

4. Add 1 mL of Binding Buffer and suspend the beads by vortexing.

5. Add 50 μL of Coupling Reagent and suspend the beads by vortexing.

6. Keep rotating the tube with Tube rotator for 10 minutes at room temperature.

7. Add 100 μg of antibody (100μL, if antibody was diluted to 1 mg/mL) and suspend the beads by vortexing.

8. Keep rotating the tube with Tube rotator for 3 hours at room temperature.

9. Remove the supernatant as in step 2.

10. Wash the beads using 1 mL of Washing Buffer and suspend the beads by vortexing.

11. Remove the supernatant as in step 2.

12. Repeat steps 10 & 11 for a total of 3 times.

13. Suspend the beads with a desired buffer suitable for downstream applications and store at 2-8 degrees C until needed.