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肽-N4-N-乙酰基-β-D-葡萄糖胺基天門冬酰胺酶F(PNGaseF)活性蛋白

Active Peptide-N4-N-Acetyl-Beta-D-Glucosaminyl Asparagine Amidase F (PNGaseF)

Ngl; PNG; PNGase F; Glycopeptide N-glycosidase; N-glycanase

  • 肽-N4-N-乙?;?β-D-葡萄糖胺基天門冬酰胺酶F(PNGaseF)活性蛋白 產(chǎn)品包裝(模擬)
  • 肽-N4-N-乙酰基-β-D-葡萄糖胺基天門冬酰胺酶F(PNGaseF)活性蛋白 產(chǎn)品包裝(模擬)
  • APX267Ge01.jpg Figure. SDS-PAGE
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

活性實(shí)驗(yàn)

PNGase F (Peptide-N-glycosidase F) is a kind of enzymes for the deglycosylation of glycoproteins. The enzyme releases asparagine-linked oligosaccharides from glycoproteins and glycopeptides by hydrolyzing the amide of the asparagine (Asn) side chain. Thus, the activity of recombinant PNGase F measured by deglycosylating Interferon Gamma (IFNg) under denatured conditions. Prepare 10×denaturing buffer(5% SDS,400mM DTT), dilute IFNg to 1 μg/μl by 1×denaturing buffer,then heat the solution to 100℃for 10 minutes to denature the glycoprotein. Cool to room temperature and microcentrifuge briefly. Dobule dilution of recombinant PNGase F by assay buffer(50mmol/L Tris(pH7.4),1% NP-40), add 10 μl denatured IFNg to 10 μl different concentration of recombinant PNGase F, incubate reaction mixture at 37℃ for 1 hour. Stop the reaction by heating to 100℃ for 5 minutes, assess deglycosylation by SDS-PAGE. In the 10 μl reaction system, the amount of PNGase F needed to remove more than 95% carbohydrates from 10 μg denatured IFNg in 1 hour at 37℃ was defined as a unit. In this procedure, one unit is equal to 2.5ng recombinant PNGase F . The results are shown in Figure 1.

用法

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

儲(chǔ)存

避免反復(fù)凍融。2-8°C不超過一個(gè)月,-80°C不超過12個(gè)月。

穩(wěn)定性

熱穩(wěn)定性以損失率顯示。損失率是由加速降解試驗(yàn)決定,具體方法如下:在37°C孵育48小時(shí),沒有顯著的降解或者沉淀產(chǎn)生。保質(zhì)期內(nèi),在適當(dāng)?shù)臈l件下存儲(chǔ),損失率低于5%。

相關(guān)產(chǎn)品

編號(hào) 適用物種:Pan-species (General,通用) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
RPX267Ge01 肽-N4-N-乙酰基-β-D-葡萄糖胺基天門冬酰胺酶F(PNGaseF)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
APX267Ge01 肽-N4-N-乙?;?β-D-葡萄糖胺基天門冬酰胺酶F(PNGaseF)活性蛋白 Cell?culture;?Activity?Assays.
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