As the CH2Cl groups onto the MS will directly react with available NH2 groups from the proteins, our modified Microspheres are classified as pre-activated product.
No pretreatment steps required.

Due to the high reactivity of these CH2Cl groups, they will dehydrohalogenate in an aqueous suspension over time, and therefore have a limited shelf life after synthesis.
Once coupled, their stability matches that of any other ligand coated Microspheres.

1°) Washing twice MS at 10 % (100mg/ml) with washing/coupling buffer (1 ml in 10 ml). Buffer :
Phosphate Buffered Saline (PBS) pH 7.5.

2°) Resuspend in 5 ml of wash/coupling buffer, (vortex, ultrasound or rolling).

3°) Dissolve protein (1-10 fold excess of calculated monolayer) in 5 ml wash/coupling buffer.
Combine Microspheres suspension and protein solution. Concentration of MS suspension is
now 10 mg/ml.

4°) React at room temperature for 3-4 hours with constant mixing.

5°) Wash, resuspend in 10 ml of quenching solution, and mix gently for 30 minutes (at room
temperature). Quenching solution with primary amine source: 30-40 mM (hydroxylamine,
ethanolamine or glycine, etc) with 0.05-1% (w/v) blocking molecule (BSA, Casein, Pepticase,
Tween 20, Triton X-100, PEG, Sera, or IgG ).

6°) Wash, and resuspend in storage buffer to desired storage concentration : often 10 mg/ml.
Storage buffer: pH 7-7.5 with 0.01-0.1% (w/v) blocking molecule and NaN3 (0.09%).

7°) Store at 4°c until used.

We supply CH2Cl-modified microspheres packaged at 10% in water.