Sulfo-MBS

The generalized protocol for performing a multi-step conjugation reaction with MBS or sulfo-MBS is similar to that described for SMCC (Section 1.3, this chapter). Specific examples may be found in the cited references. 

Dissolve 5 mg of MBS in a small amount of dimethylformamide (about 0.75 mL) or for Sulfo-MBS, dissolve in a small amount of sterile water. 

MBS Sulfo-MBS version of this reagent is water-soluble and, therefore, preferable (Pierce). 

If, during MBS coupling, the DMF concentration exceeds 30% the KLH will come out of solution. KLH concentrations in excess of 20 mg/mL will also lead to insolubility. Use the Sulfo-MBS derivative if possible. 

---