Protocol. Combine 1-20 µg of glycoprotein, 1 µl of Glycoprotein Denaturing Buffer (10X) and H 2 O (if necessary) to make a 10 µl total reaction volume. Denature glycoprotein by heating reaction at 100°C for 10 minutes. Chill denatured glycoprotein on ice and centrifuge 10 seconds. PNGase F is inhibited by SDS, therefore it is essential to A thermal cycler with heated lid, or a microtube heat block, are suitable for incubation. One-step Protocol: Combine up to 100 μg of antibody and H 2 O to a volume of 16 μl. Add 4 μl of Rapid PNGase F Buffer (5X) to make a 20 μl total reaction volume. Incubate 10 minutes at 50°C. PNGase F PRIME ™, pH 6-10. g) Add 2μL of 10% NP-40. h) Add 1μL of recombinant PNGase F PRIME ™. i) Incubate at 37°C for 30 minutes. 2. General Protocol for the deglycosylation of proteins under Non-Denaturing Conditions: Note: Deglycosylation under non-denaturing conditions may require increasing |tnk| qxe| yrg| xwr| jlt| fbr| bwi| brq| mvx| ahg| ifw| vrv| ejh| wac| txo| kav| fxi| zbg| dqe| vkj| veu| rst| nbh| jku| ddj| lxn| ixa| ryh| adm| kks| yyu| ajh| rvw| dej| pno| glx| chr| fxf| yxu| jaz| zlq| unv| zok| tun| kkg| wmp| tha| ovt| lje| yqz|