Can I reuse the immobilized digestion enzymes in the Thermo Scientific IgG Fragmentation kit?
No. For best results, use the immobilized enzyme once and discard after use.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
How long does IgG digestion take using the Thermo Scientific IgG Fragmentation kit?
The digestion time is a function of the IgG concentration and source. For rabbit, mouse and human IgG, optimized digestion times are given in the instructions for each kit. Digestion times listed in the product instructions result in 80-90% digestion, although efficiency varies depending on IgG preparation and source.
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In the Thermo Scientific IgG Fragmentation kit, when digesting an IgG that does not bind to Protein A, how do I purify the Fab and F(abÁ)2 fragments?
If the IgG is a species or subclass that does not bind to Protein A, researchers can immobilize an antibody specific for the desired fragment. For example, to purify digested rat IgG2a, use the Aminolink Plus Immobilization Kit (Cat. No. 44894) to immobilize goat anti-rat Fc antibody. Our Tech Tip #34 and other web resources list IgG binding properties of Protein A. (Because Protein G and Protein A/G bind Fab fragments, they are not effective for this application.)
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Can I purchase the immobilized enzymes (proteases) from the Thermo Scientific IgG Fragmentation kit separately?
Yes. The three immobilized enzymes are available in 5 mL package sizes as shown below:
Immobilized Ficin, Cat. No. 44881
Immobilized Papain, Cat. No. 20341
Immobilized Pepsin, Cat. No. 20343
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
How many Thermo Scientific IgG Fragmentation kits are available and which enzymes are included?
There are six Thermo Scientific IgG fragmentation kits:
Fab Preparation Kit (Papain), Cat. No. 44985
Fab Micro Preparation Kit (Papain), Cat. No. 44685
F(abÁ)2 Preparation Kit (Pepsin), Cat. No. 44988
F(abÁ)2 Micro Preparation Kit, (Pepsin), Cat. No. 44688
Mouse IgG1 Fab and F(abÁ)2 Preparation Kit (Ficin), Cat. No. 44980
Mouse IgG1 Fab and F(abÁ)2 Micro Preparation Kit (Ficin), Cat. No. 44680
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.