FragIT™ Smart Enzymes

FabRICATOR digests IgG at a specific site below the hinge region, generating a homogenous pool of F(ab')2 and Fc/2 fragments. The IgG is digested within 30 minutes, and there is no risk of overdigestion if the incubation time is prolonged. Since FabRICATOR digests IgG under physiological reaction conditions, the immunoreactivity is preserved.

FragIT is a resin with FabRICATOR enzyme covalently coupled to agarose beads for generation of F(ab’)2 and Fc/2 fragments without contaminating the final preparation with enzyme. The workflow is simple – IgG is incubated with the FragIT resin and fragments are easily collected by a centrifugation step. The spin columns are available for digestion of up to 0.5 mg (Microspin), 10 mg (Midispin) or 100 mg (Maxispin) IgG. The smallest columns are available in three different pack sizes, with 2, 5 or 10 columns per pack.

FragIT digests all subclasses of human, and some classes of monkey, rabbit and sheep IgG. It has limited activity on mouse IgG2a and IgG3 – for digestion of these antibody species, we recommend using FragIT Z .

Storage and Stability

The FragIT spin columns are supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the products!

Buffers

FragIT can be used with all commonly used buffers with pH ranging from 6.0 to 8.0. Optimization may be required. Note that the FabRICATOR enzyme is irreversibly inactivated below pH 5.0.

Unit Definition

One spin column contains sufficient FabRICATOR-coupled agarose beads to digest 0.5 mg (Microspin), 10 mg (Midispin), or 100 mg (Maxispin) IgG.