The Fab & F(ab)2 Fragmentation of Mouse IgG1 kit is designed for the generation and isolation of Fab and F(ab)2 fragments from mouse IgG1 molecules.
The kit utilizes our Immobilized Ficin resin. Ficin (or Ficain) (~25,000Da) is a cysteine protease enzyme (EC 3.4.22.3) isolated from fig latex is that has the endopeptidase activity to cleave immunoglobulin G molecules in the hinge reason. Ficin has an effective range of pH4-9.5 with an optimal pH of 6.5 and cleaves bonds that involve uncharged or aromatic amino acids.
Ficin is typically used to cleave mouse IgG1 as this are difficult to cleave with papain and pepsin. In the presence of 1-4mM or 10-20mM cysteine, ficin generates F(ab′)2 and Fab fragments respectively. Immobilized Ficin is a convenient reagent for producing Fab and F(ab′)2 fragments as it avoids the need to remove the ficin enzyme after digestion.
Following ficin digestion the fragments are separated from undigested IgG and the large Fc fragments with the supplied Protein A (Cat. # 786-276, 786-277) or Protein G (Cat. # 786-1276) Spin Column. The Protein A Resin or Protein G Resin binds the IgG and Fc molecules and the Fab or 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 fragmentation.
The Fab & F(ab)2 Fragmentation of Mouse IgG1 kit is optimized for mouse IgG1, using 0.25-4mg/ 0.5ml sample.
The Fab & F(ab)2 Fragmentation of Mouse IgG1 Micro kit is optimized for mouse IgG1, using 25-250µg/ 125µl sample.
Features
- Immobilized Ficin ensures no enzyme contamination of digestions
- Optimized for mouse IgG1
- Results in ready-to-use Fab or 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 preparation and purification of Fab or F(ab)2 fragments from mouse IgG1 antibody molecules.