Genovis » FragIT kit

Name: FragIT kit
SKU: 9475
Price: 380.00 EUR
Availability: InStock

Description

FragIT kit contains immobilized FabRICATOR (IdeS) enzyme on agarose beads in spin columns and CaptureSelect^TM for affinity capture of Fc fragments. FragIT digests antibodies or Fc-fusion proteins at one single amino acid position below the hinge region generating Fc and F(ab’)2 fragments. CaptureSelect^TM spin columns capture Fc fragments for purification of F(ab’)2 and Fc fragments (other CaptureSelect^TM resin needed for rabbit IgG, please inquire). FragIT kit provides digestion and purification of 0.5 – 100 mg of antibody or Fc-fusion protein without residual enzyme in the final preparation.

Detailed Information

Application	Antibody Fragmentation, Antibody Characterization, Purify F(ab')2 and Fc Fragments
Product Group	FabRICATOR
Source/Origin	Cloned from S. pyogenes, Produced in E. coli (His-tagged), 37.7 kDa
Product Composition	Immobilized on agarose beads (20% EtOH), CaptureSelect Fc columns included
Digestion Site	Below Hinge
Specificity	CPAPELLG / GPSVF (below the hinge)
Substrates	Human IgG1, Human IgG2, Human IgG4, Monkey IgG, Rat IgG, Sheep IgG
Reaction Conditions	30 minute reaction at 37°C according to protocol specifications
Digestion Buffers	PBS (pH 7.4)
Buffer Compatibility	Other commonly used buffers at physiological pH (6.0 – 8.0) and ionic strength can also be used. Optimization may then be required.
Inhibitors	Iodoacetate 20 mM, Iodoacetamide 1 mM, pH < 5.1
Unit Definition	0.1 ml settled resin digests ≥ 95% of 0.5 mg Human IgG1

FAQs

Can I re-use FragIT columns?

We can only guarantee optimal digestion for one time use. However, depending on antibody the columns can be reused but we do not have a proper sanitization protocol. We don’t recommend that you run different antibodies on the same column as we cannot guarantee that there will be no carry-over. If desired to reuse the column store in 10-20% ethanol at +4-8°C.

Can I use protein A or MabSelectSure (GE) for removal of Fc-fragments?

To use protein A or MabSelect Sure (GE) for removal of Fc as a platform after digestion with FabRICATOR or FragIT is problematic. Therefore we have chosen CaptureSelect in the FragIT Kit for removal of Fc fragments. CaptureSelect is much more specific for Fc. Both protein A and MabSelect Sure (GE) may have affinity for Fab and the degree of affinity is antibody dependent.We have many customer reports on this subject and they usually comes back to us for the CaptureSelect columns for the above reason.

How do I release Fc fragments from CaptureSelect column after isolation of F(ab’)2 fragments.

You can indeed elute the Fc fragments from the CaptureSelect column using a low pH buffer commonly used for example during protein A/G purification of antibodies. We have successfully used 0.1 M glycine buffer pH 2.6-2.8 to release the Fc fragments. Be sure to adjust pH back to normal using a 1 M Tris-HCl buffer or similar directly after isolation of the Fc fragments from the CaptureSelect column.

Where does FabRICATOR cleave the IgG molecule?

FabRICATOR® cleave the IgG molecule in one specific site in the lower hinge region of the anitbody leaving a F(ab’)2 fragment and a Fc-fragment.
Human IgG1: HTCPPCPAPELLG / GPSVF
Human IgG2: VECPPCPAPP_VA / GPSVF
Human IgG3: PPCPRCPAPELLG / GPSVF
Human IgG4: PHAHHAQAPEFLG / GPSVF

FragIT kit, Digest and Purify up to 0.5 mg quantity	FragIT kit, Digest and Purify up to 0.5 mg (A2-FR2-005)	€400.00
FragIT kit, Digest and Purify up to 5 x 0.5 mg quantity	FragIT kit, Digest and Purify up to 5 x 0.5 mg (A2-FR2-025)	€1,070.00
FragIT kit Midispin, Digest and Purify 1-10 mg quantity	FragIT kit Midispin, Digest and Purify 1-10 mg (A2-FR2-100)	€1,340.00
FragIT kit Maxispin, Digest and Purify 10-100 mg quantity	FragIT kit Maxispin, Digest and Purify 10-100 mg (A2-FR2-1000)	€4,010.00

FragIT kit

Description

Detailed Information

FAQs

Can I re-use FragIT columns?

Can I use protein A or MabSelectSure (GE) for removal of Fc-fragments?

How do I release Fc fragments from CaptureSelect column after isolation of F(ab’)2 fragments.

Where does FabRICATOR cleave the IgG molecule?

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