Frequently Asked Questions

Got a question? There’s a good chance we’ve answered it here. Browse our FAQs or use the filters below to find what you're looking for. Couldn't find an answer to your question here? Contact us!

Find your question

Can't find an answer to your question? Contact us!

Which buffers are compatible with IgASAP Sub1?

The enzyme is active at pH 6.5 to 8.0, tolerates up to 300 mM NaCl, and does not require reducing conditions or co-factors for activity. PBS and TBS buffers are both compatible. Optimization may be required if buffers other than the recommended are used.

Learn more about IgASAP™

Can the IgASAP Sub1 digestion be performed at lower temperatures?

The optimal activity is at 37°C, but the digestion can also be performed at room temperature using a prolonged incubation time.

Learn more about IgASAP™

What enzyme should be selected for the digestion of IgA1?

IgASAP Sub1 specifically targets IgA1 and exhibits a faster reaction time (1 hour) compared to IgASAP Sub1+2 (16-18 hours). The cleavage site of IgASAP Sub1+2 is located six amino acids above that of IgASAP Sub1, resulting in the absence of potentially O-glycosylated residues on the Fab fragments. This characteristic can be advantageous in certain cases.

Learn more about IgASAP™

Is there an enzyme for digestion of IgA2-only?

There is no known enzyme that targets IgA2-only since the amino acids in the hinge regions of IgA2 is present also in IgA1. However, IgASAP Sub1 will digest IgA1-only.

Learn more about IgASAP™

Is the antibody in Anti-FabRICATOR Monoclonal Biotin Conjugated (A3-AF5-005) from the same clone as Anti-FabRICATOR Monoclonal (A3-AF4-010)?

No, the monoclonal for biotinylation is produced by another clone of mouse hybridoma.

Learn more about Anti-FabRICATOR®

Are there any mutated IgG formats that FabRICATOR Xtra LALA cannot digest?

Yes, of the formats we have specifically tested, hIgG1-LALAPG (L234A, L235A, P329G), hIgG1-STR (L234S, L235T, G236R) and hIgG1-L234F, L235E, D265A could not be digested to satisfactory levels using FabRICATOR Xtra LALA. Please refer to footnote 4 in the instructions for use to determine whether your antibody is likely to be suitable for FabRICATOR Xtra LALA digestion.

Learn more about FabRICATOR® Xtra

Can I release Fc fragments from the CaptureSelect column after isolating the F(ab’)2 fragments?

It is possible to elute the Fc fragments from the CaptureSelect column using a low pH buffer, for example a 0.1M Glycine buffer pH=3.0. The pH must immediately after elution be adjusted to neutral pH by adding 0.1x elution volume of 1M Tris pH=8.0.

Learn more about FabRICATOR®

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

Genovis has chosen CaptureSelect in the FabRICATOR Fab2 Kit for removal of Fc fragments. CaptureSelect has a very high specific affinity for the Fc. Both protein A and MabSelect Sure (GE) may have affinity for the Fab which may result in a lower recovery of the generated F(ab’)2 fragments and the degree of affinity is antibody dependent. We have many customer reports on this subject, and they usually come back to us for the CaptureSelect columns for the above reason.

Learn more about FabRICATOR®

Does FabRICATOR introduce modifications?

FabRICATOR is a cysteine protease. It should not result in any other modifications than the digestion of the peptide bond. Digestion will be between the two Glycines (…CPAPELLG / GPSVF…) and the C-terminus will regain the carboxyl group – that is addition of one OH-group and the N-terminal amino group will be regained (addition of one H).

In summary, R1-CO-NH-R2 + H2O –> R1-COOH + NH2-R2.

Learn more about FabRICATOR®

Can I use FabRICATOR Fab2 Kit for generation and purification of fragments from Rabbit IgG?

FabRICATOR Fab2 Kit digests rabbit IgG but to capture the Fc fragments in the affinity step you need another resin than the CaptureSelect™ Fc normally included in the kit. Please contact the Genovis team if you are interested in processing rabbit IgG and we will provide columns with resin compatible with rabbit Fc in the FabRICATOR Fab2 Kit.

Learn more about FabRICATOR®