G-Biosciences' Mouse IgG₁ Fab & F(ab)₂ Preparation Kit is designed for the generation and isolation of Fab and F(ab)₂ fragments from mouse IgG₁ molecules

Ficin (~25,000Da) is a cysteine protease enzyme 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 1-4 mM or 10-20 mM 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 Spin Column. The Protein A 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 0.25-4 mg/ 0.5 mL sample. The Fab & F(ab)₂ Fragmentation of Mouse IgG₁ Micro kit is optimized for mouse IgG₁, using 25-250 µg/ 125µL sample.

The kit utilizes our Immobilized Ficin resin.