The Gly-X™ N-Glycan Rapid Release and Labeling with APTS Express™ kit utilizes a novel in-solution enzymatic protein deglycosylation followed by Express labeling of released N-glycans with 8-aminopyrene-1,3,6-trisulfonic acid (APTS). With deglycosylation carried out in solution, the method is simple, rapid and suitable for automation. After a simple clean-up step, the APTS-labeled glycan samples are ready for analysis by CE, CE-MS, and other methods.
Sample prep time: ~2.5 hours.
Other benefits include:
Equipment and reagents provided by user:
Size: 1 kit (24-ct)
Product Code: GX24-APTS
Q. Can I use more than the recommended upper limit of 40 µg protein per reaction?
A. Maybe, although it will depend on the protein. You would have to test to ensure that labeled glycans from > 40 µg of your protein can be successfully loaded onto the Gly-X Cleanup Matrix without blockage.
Q. My samples are not loading completely onto the Gly-X Cleanup Matrix. What is happening?
A. This may be caused by the nature of your protein sample, or by matrix effects caused by the composition of your formulation buffer. This can be addressed by using less protein per reaction, or by buffer exchanging your protein prior to starting the Gly-X protocol.
Q: Can I use Eppendorf microtubes for the Denaturation and Digestion Steps, rather than a PCR plate?
A: Yes, leave the tubes open for the heating steps, and ensure that material does not get in the lid during mixing.
Q: Is it possible to use a plate shaker/heater such as the Eppendorf Thermo Mixer C formixing and heating during the Denaturation and Digestion protocols?
A: Yes, this is possible. Mix rate should be high enough to vortex material in wells.
Q: What is the Finishing Reagent for prior to the APTS labeling step?
A: The Finishing Reagent is a weak acid, and drives conversion of the glycosylamine (-NH2) to a glycan with a free reducing end (-OH) prior to APTS labeling by reductive amination.
Product Safety Documentation for GX24-APTS:
|WS0343||Gly-X N-Glycanase (PNGase F)|
|WS0344||Gly-X Digestion Buffer|
|WS0360||Gly-X APTS Reductant Solution|
|WS0380||Gly-X APTS Solution|
|WS0381||Gly-X APTS Catalyst|
|WS0385||Gly-X APTS Finishing Reagent|
This kit contains multiple modules, each with their own Certificate of Analysis, which may be searched for & accessed here.