Fab Preparation kits, Pierce™
Pierce™ Fab preparation kits use immobilised papain protease to digest human or mouse IgG antibodies to make separate Fab and Fc fragments and subsequently to purify the Fab using Protein A agarose.
- High capacity – use the standard kit for fragmenting 0,25 to 4 mg IgG or the micro kit for 25 to 250 µg IgG
- Immobilised Papain (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
- Digestion and final recovery of purified Fab fragments occurs in neutral pH sodium phosphate buffer, suitable for storage or immediate use in typical applications
- Complete kits include all reagents needed to prepare and purify antibody fragments
- Spin format reduces sample processing time
- Protocols are included for multiple species and IgG subclasses, as well as sample size and concentration
The kit procedure is optimised for human, mouse and rabbit IgG. Papain based digestion is effective for certain other species and subclasses of IgG (eg. goat and pig), although the purification step requires that the antibody effectively binds to Protein A.
: Each standard kit includes immobilised papain, 1,25 ml; cysteine-HCl, 1 mg; Fab digestion buffer, 120 ml; NAb™ Protein A Plus spin columns, 1×1 ml; BupH™ PBS, 2 packs (makes 1L); IgG elution buffer, 120 ml; spin columns, 10×0,8 ml; microcentrifuge tubes, 30×2 ml; and Zeba™ desalting columns, 7K MWCO, 10×2 ml. Each micro kit includes immobilised papain, 0,5 ml; cysteine-HCl, 1 mg; Fab digestion buffer, 55 ml; NAb™ Protein A Plus spin columns, 2×0,2 ml; BupH™ PBS, 2 packs (makes 1L); IgG elution buffer, 50 ml; spin columns, 10×0,8 ml; microcentrifuge tubes, 30×2 ml; and Zeba™ desalting columns, 7K MWCO, 10×0,5 ml.