Product Description
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| SDS | USER MANUAL CM51407 |
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 molecular weight, and experience lower nonspecific binding to cell surfaces. Outside of antibody-enzyme conjugates, Fab fragments are also 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. Then a purification step to remove Fc fragment and obtain the pure Fab.
This kit provides materials to fragment 0.25-3 mg of mouse IgG1 antibodies with the option to choose from one (CM51407x1) or three (CM51407x3) reactions.
Key Features of this PerKit™
- Easy preparation: 1 day preparation with <2 hrs hands-on time
- All reagents and supplies included for preparation and purification
- 40-60% recovery of 80-90% of pure Fab by SEC (size-exclusion chromatography) HPLC Analysis
- Options to choose tailored services at CellMosaic® after conjugation
After conjugation: you can choose to send your sample to CellMosaic® for HPLC analysis.
Typical Kit Performance Data
Figure 1. SEC HPLC analysis of a mouse IgG1, crude digestion mixture of the IgG1, and purified Fab (UV at 220 nm)
