The Protein Deglycosylation Mix contains PNGase F, O-Glycosidase, Neuraminidase (sialidase), β1-4 Galactosidase, and β-N-Acetylglucosaminidase. It is used to simultaneously remove N-glycans and some O-glycans8 . Finally, the Deglycosylation Mix was supplemented with a mixture of other exo...
Protein deglycosylation mix kit (Promega) was used to detect glycosylation of EMP1. Cell surface proteins were isolated and collected by Mem-PERTM Plus Membrane Protein Extraction Kit (Thermo Fisher Scientific). For immunoprecipitation, supernatants were incubated with primary antibody (1:100 dilution)...
Deglycosylation Of each SAPN, 15.5 μg was brought up to 49 μL in the respective refolding buffer. One microliter of Protein Deglycosylation Mix II (New England Biosciences, Ipswich, MA) was added to the samples. Reactions were held at 25 °C for 30 minutes and proteins were identified on...
Protein Deglycosylation kit – enzymes mix NEB P6044S StepOnePlus – Real-Time PCR System Applied Biosystems 4376600 Deposited data Results related to Figure 1A - Healthy Blood Donors (HBD) https://data.mendeley.com/datasets/rc74sdgksk/1 Additional supplemental items are available from Mendeley Da...
or toward the supply of precursors for cell wall polysaccharide synthesis and protein glycosylation27(Fig.5). HL suppressed the transcripts of the cellulose biosynthesis gene (i.e.,g7046) (Supplementary Fig.10d) and a gene involved in protein deglycosylation (i.e.,g9910) (Supplementary Fig.10...
For visualization of deglycosylated proteins on SDS-PAGE, the isolated proteins were incubated with a Protein Deglycosylation Mix (New England Biolabs, Mississauga, ON) according to the manufacturer’s instructions. Briefly, 9 μg of purified proteins were incubated with 40 MM DTT and 0.5% SDS at...
Msp1 was shown to stain positive with periodic-acid Schiff staining, to be susceptible to chemical deglycosylation, and to bind with the mannose-specific Concanavalin A (ConA) lectin. Recombinant expression in Escherichia coli resulted in a significant reduction in molecular mass, loss of ConA ...
Upon deglycosylation with PNGase F, C1 became detectable as a band of approx. 16 kDa. We could detect C1 irrespectively of the ADAM10 status of neurons (Figure 5A; complete Western blot of PNGase F digestion is shown in Additional file 1B). In fact, C1 signal strength was stronger in ...
For deglycosylation antibodies were added to the column and incubated at room temperature for 30 min with end-over-end mixing. Afterwards, antibodies were collected by centrifugation at 1,000g for 1 min. To maximize the recovery, 100 µl TBS was added to the column and centrifuged ...
Deglycosylation of the recombinant T97 restored the proteins cytotoxicity suggesting that glyco- sylation are responsible for this inhibitory role. The physiological function of the granule contained cytotoxic ribonuclease Eosinophils contain a large amount of ECP but the ques- tion is why? What is the...