In the preparation of antibody-enzyme conjugates there are a few advantages of using antibody fragments rather than intact antibodies. For example, antibody fragments lacking an Fc region have less interference with various Fc binding proteins, can easy penetrate cell membranes due to their lower molecule weight, and experience lower nonspecific binding to cell surfaces. Fab fragments are useful in immuno-histochemical studies.

CellMosaic has designed this kit for Fab preparation from mouse IgG1 antibody. The procedure we use involves cleaving the IgG molecule in the hinge region above the interchain disulfides using soluble Ficin to create two identical Fab portions and one intact Fc fragment, and then purification to afford the pure Fab.

This kit provides materials to fragment 0.25-3 mg of mouse IgG1 antibodies with the option to choose from one or three reactions.

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Key Features of this PerKit™

• Easy preparation: 1 day preparation and <2 h hands-on time
• All reagents and supplies included for preparation and purification
• 40~60% recovery and 80-90% of pure Fab by SEC (size-exclusion chromatography)
• Options to choose tailored services at CellMosaic after conjugation

After conjugation: you can choose to send your sample to CellMosaic for HPLC analysis.

Examples

Figure 1. SEC analysis of a mouse IgG1, crude digestion mixture of the IgG1, and the purified Fab (UV at 220 nm)