Lysates of patient- and donor-derived LCLs were treated with enzymatic de-glycosylation (i.e. PNGase F, Endo-O-Glycosidase, α-2[3, 6, 8, 9]-neuraminidase, position-specific β-1–4-galactosidase, and β-N-acetylglucosaminidase; EDEGLY-Kit, Sigma).
Edegly kit
Manufactured by Merck Group
The EDEGLY kit is a laboratory equipment product manufactured by Merck Group. It is designed for use in scientific research and analysis applications. The kit includes essential components necessary for conducting experiments and analytical procedures. The core function of the EDEGLY kit is to facilitate specific laboratory tasks, but no further details on its intended use can be provided in an unbiased and factual manner.
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Lab products found in correlation
4 protocols using edegly kit
1
Enzymatic De-glycosylation of Cell Lysates
2
Deglycosylation of Ebola GP Effects
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For GP deglycosylation, supernatants containing EBOV GPs were treated using EDEGLY kit (Sigma) for 48 h according to the manufacturer's instructions. Shed GP mixed with deglycosylation enzymes pretreated at 80°C for 30 min were used as a control. Monocyte-derived DCs and macrophages were treated with deglycosylated shed GP samples for 24 hours and culture supernatants were assayed for the presence of TNFα using a Multiplex cytometric bead array (Bio-Rad) in Luminex MAGPIX.
3
Protein Deglycosylation Protocol
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N-deglycosylation: 250 ng of purified proteins were denatured at 95 °C for 10 min in 10 mM potassium phosphate and 0.2% SDS. The samples were then incubated overnight at 37 °C in a buffer (10 mM potassium phosphate, pH8, 10 mM EDTA, 0.5% Triton X-100, 0.2% SDS, 1% β-mercaptoethanol) containing 0.1–1 U of PNGase F (Roche Applied Science).
O-deglycosylation: The O-deglycosylation was performed with the EDEGLY kit (Sigma-Aldrich).
250 ng of purified proteins were denatured at 100 °C for 5 min in 2.5 μl of denaturation solution (Sigma-Aldrich) and 10 μl of reaction buffer 5X. Samples were incubated overnight at 37 °C with 1 μl of each enzyme (PNGase F, O-Glycosidase, α-(2→3,6,8,9)-Neuraminidase, β-N-Acetylglucosaminidase, β-(1→4)-Galactosidase).
O-deglycosylation: The O-deglycosylation was performed with the EDEGLY kit (Sigma-Aldrich).
250 ng of purified proteins were denatured at 100 °C for 5 min in 2.5 μl of denaturation solution (Sigma-Aldrich) and 10 μl of reaction buffer 5X. Samples were incubated overnight at 37 °C with 1 μl of each enzyme (PNGase F, O-Glycosidase, α-(2→3,6,8,9)-Neuraminidase, β-N-Acetylglucosaminidase, β-(1→4)-Galactosidase).
4
Deglycosylation of Purified FST Isoforms
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A 1 mg aliquot of each of the purified FST isoforms was treated with a mixture of five different deglycosylation enzymes (E-DEGLY kit; Sigma) according to the manufacturer's instructions. Treated vs non-treated samples were run on SDS-PAGE and stained with silver nitrate according to the method described by Morrissey (1981) (link).
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