The F(ab)2 Fragmentation kits are designed for the generation and isolation of F(ab)2 fragments from IgG molecules.
The kit utilizes our Immobilized Pepsin resin. Pepsin is a proteolytic enzyme that is routinely used for the generation of F(ab)2 fragments from immunoglobulin G (IgG). The pepsin has the ability to cleave the heavy chains near the hinge region. One or more of the disulfide bonds that join the heavy chains in the hinge region are preserved, so the two Fab regions of the antibody remain joined together, yielding a divalent molecule (containing two antibody binding sites), hence the designation F(ab)2. The light chains remain intact and attached to the heavy chain, whereas the Fc fragment is digested into small peptides. The Immobilized Pepsin offers the distinct advantage of eliminating enzyme contamination of the F(ab)2 fragments.
Following pepsin digestion the F(ab)2 fragments are separated from undigested IgG and the large Fc fragments with the supplied Protein A Spin Column or the Protein G Spin Column. The Protein A Resin or Protein G binds the IgG and Fc molecules and the F(ab)2 are rapidly collected due to the spin-format design.
In addition, SpinOUT™ GT-600 desalting columns are supplied to ensure the initial antibody sample is in the optimal condition for F(ab)2 Fragmentation.
The F(ab)2 Fragmentation kit is optimized for mouse, rabbit and human IgG, using 0.25-4mg/ 0.5ml sample.
The F(ab)2 Fragmentation (Micro) kit is optimized for mouse, rabbit and human IgG, using 25-250µg/ 0.125ml sample.
Features
- Immobilized Pepsin ensures no enzyme contamination of digestions
- Optimized for human, mouse* and rabbit IgG
- Results in ready-to-use F(ab)2 fragments, with enhanced yield and purity
- Spin format for rapid purification
- Contains all essential reagents
- Two convenient kit sizes available
Applications
- For the generation of F(ab)2 fragments from IgG antibody molecules