|SDS||USER MANUAL CM51408|
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 easily penetrate cell membranes due to their lower molecule weight, and experience a lower nonspecific binding to cell surfaces. F(ab')2 and Fab' fragments are useful in immuno-histochemical studies.
CellMosaic has designed this kit for F(ab')2 preparation from mouse IgG1 antibody. The procedure we use involves cleaving the IgG molecules below the disulfide groups in the hinge region using Ficin to create a bivalent fragment and degraded smaller peptide fragments, and then purify to obtain F(ab')2. F(ab')2 can be further reduced to produce Fab' fragment. If you need Fab', contact CellMosaic for a custom synthesis.
This kit provides materials to fragment 0.25-3 mg of mouse IgG1 with the option to choose from one or three reactions.
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 80-95% of pure F(ab')2 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.
Figure 1. SEC analysis of a mouse IgG, crude digestion mixture of the IgG, and the purified F(ab')2 (UV at 220 nm)