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PNGase F

规格或纯度: Specific Activity >25 U/mg;Activity 5 U/ml

有货

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库存信息

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货号 (SKU)	包装规格	是否现货	价格	数量
P420186-20μl	20μl	期货
P420186-60μl	60μl	期货

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基本描述

规格或纯度	Specific Activity >25 U/mg;Activity 5 U/ml
英文名称	PNGase F
英文别名	Peptide-N4-(acetyl-?-glucosaminyl)-asparagine amidase N-Glycosidase F
来源	Elizabethkingia meningosepticum was (Chyrseobacterium/Flavobacterium men.)
储存温度	2-8°C储存
运输条件	冰袋运输
产品介绍	Product Description PNGase F cleaves N-linked (asparagine-linked) oligosaccharides from glycoproteins. The enzyme deaminates asparagine to aspartic acid, leaving the oligosaccharides intact. PNGase F will not remove oligosaccharides containing Alpha-(1,3)-linked core fucose commonly found on plant glycoproteins; for this purpose, use peptide N-glycosidase A. Denaturation increases the rate of cleavage. Most native proteins can still be completely N-deglycosylated but incubation time may need to be increased. The enzyme will remain fully active under reaction conditions (37?C) for at least 96 hours. There are a number of alternative enzymes which can be used to remove N-glycans, most especially the Endo F family of enzymes and Endo H. These enzymes cleave between the two N-acetylglucosamine residues in the core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. This enhances the solubility, keeping proteins in solution that precipitate after deglycosylation with PNGase F which removes the oligosaccharide intact. Endo F1cleaves high mannose and some hybrid type N-glycans. Endo F2 will removes biantennary and high mannose (at a 40X reduced rate). Endo F3releases of triantennarry and fucosylated biantennary N-glycans. Endo H removes hybrid or high mannose glycans. Contents? PNGase F in 20 mM Tris-HCl, pH 7.5 Included with 20 μL and 60 μL pack sizes: 5x Reaction Buffer 7.5 – 250 mM sodium phosphate, pH 7.5 Denaturation Solution – 2% SDS, 1 M Beta-mercaptoethanol Triton X-100 – 15% solution Specific Activity?>25 U/mg Activity?5 U/ml Molecular weight?35,000 daltons pH range?6-10, optimum 7.5 PNGase F Protocol 1. Add up to 200 μg of glycoprotein to an Eppendorf tube. Adjust to 35 μl final volume with de-ionized water. 2. Add 10 μl 5x Reaction Buffer 7.5 and 2.5 μl of Denaturation Solution. Heat at 100°C for 5 minutes. 3. Cool. Add 2.5 μl of Triton X-100 and mix. 4. Add 2.0 μl of enzyme to the reaction. Incubate 3 hours at 37°C. Specifictity? Cleaves all asparagine-linked complex, hybrid or high mannose oligosaccharides unless alpha(1-3) core fucosylated; asparagine must be peptide bonded at both termini, Endo F free Specific Activity Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured RNase B in 1 minute at 37°C, pH 7.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster).

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