Does IgMBRAZOR digest only human IgM?
IgM from some species of monkey may also be digested by IgMBRAZOR.
Can IgMBRAZOR and FabRICATOR be used in the same reaction?
Yes, IgMBRAZOR and FabRICATOR can be used in combination.
Can IgMBRAZOR and PNGase F be used in the same reaction?
Yes, IgMBRAZOR and PNGase F can be used in combination if the reaction is performed under non-denaturing conditions. However, for complete deglycosylation of IgM using PNGase F denaturing conditions may be required and it is then recommended to perform the IgMBRAZOR digestion prior the PNGase F deglycosylation.
Is it possible to use IgMBRAZOR with very low concentrations of IgM?
It is possible to use IgMBRAZOR with low concentrations of IgM. However, this will lead to a decreased reaction rate, so for IgM concentrations below 0.5 mg/mL it is recommended to increase the amount of added enzyme and/or increase the incubation time. There is no risk of overdigestion and the incubation time can be prolonged to overnight if necessary.
Which buffers are compatible with IgMBRAZOR?
The enzyme is active at pH 5.5 to 9.0, 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.
Is it possible to reuse the OmniGLYZOR column?
No, it is not recommended. We can only guarantee optimal performance for one-time use.
Does OmniGLYZOR remove all N-glycans?
OmniGLYZOR hydrolyzes the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides. It does not remove N-glycans with α1-3 core fucosylation.
Can OmniGLYZOR be used for released glycan analysis?
No, both N- and O-glycans are trimmed by the exoglycosidases present in OmniGLYZOR and do not longer represent the structures found on the intact glycoprotein substrate.
Does PNGase F Immobilized remove all N-linked glycans?
PNGase F Immobilized hydrolyzes the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides. It does not remove N-glycans with α1-3 core fucosylation.