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QA-Bio-甲基化板(LC-PERMET-96)是可擴(kuò)展的,可以用于1到96個(gè)樣本的數(shù)量。將未用的試劑盒組分和未用的反應(yīng)板孔存放在推薦的儲(chǔ)存條件下,以便日后進(jìn)一步的樣本準(zhǔn)備。
注意:由于對(duì)碘甲烷/甲基碘的運(yùn)輸限制,無法將這一組分隨試劑盒提供。建議從您當(dāng)?shù)氐幕瘜W(xué)品供應(yīng)商處購買純度≥99.0%(氣相色譜法)的甲基碘。甲基碘的同義詞:碘甲烷,CAS編號(hào):74-88-4。

艾美捷QA-Bio-過甲基化試劑盒(LT-PERMET-VP96)組成:
96 well Permethylation plate 1 Plate
Dimethyl sulfoxide (DMSO) 40.0 ml
Dichloromethane (DCM) 60.0 ml
96 well Balance Plate 1 Plate
96 well Plate Lid 1 Lid / Mat
QA-Bio-過甲基化試劑盒用于糖苷的甲基化處理。甲基化可以穩(wěn)定唾液酸,以便進(jìn)行MALDI質(zhì)譜分析,并有助于連接分析研究(MALDI-MS,ESI-MS)。
提高并增強(qiáng)質(zhì)譜中糖苷的電離效率
增加糖苷的疏水性能,使其能夠進(jìn)行液相色譜-質(zhì)譜(LC-MS)分析
可執(zhí)行相對(duì)定量和絕對(duì)定量
穩(wěn)定唾液酸基團(tuán)
更容易確定分支和糖苷鍵的位置
能夠在正離子模式下檢測(cè)中性糖苷和酸性糖苷
時(shí)間表
步驟1:樣品的過甲基化
a. Preparation of glycans As required
b. Addition of DMSO and sample to plate 15 minutes
c. Incubation 15 minutes
d. Addition of MeI to permethylation plate 15 minutes
e. Incubation 60 minutes
步驟2:全甲基化樣品的提取和儲(chǔ)存
a. Extraction of samples 3-4 hours
b. Drying permethylated samples 10-30 minutes
c. Addition of 70% MeOH for storage 10-20 minutes
QA-Bio-過甲基化試劑盒文獻(xiàn)參考:
A. Dell; A. J. Reaseon; Kay-Hooi Khoo; M. Panico; A. McDowell and H. R. Morris Mass Spectrometry of Carbohydrate containing Biopolymer Methods in Enzymology 230: 108-132 (1994)
I.Ciucanu; C. E. Costello Elimination of Oxidative Degradation during the per-O-Methylation of Carbohydrates J. Am. Chem. Soc. 125: 16213-16219 (2003)
I.Ciucanu Per-O-Methylation reaction of structural analysis of carbohydrates by mass spectrometry Analytica Chimica Acta 576: 147-155 (2006)
I.Ciucanu; R. Caprita Per-O-Methylation of neutral carbohydrates directly from aqueous samples for gas chromatography and mass spectrometry analysis Analytica Chimica Acta 585: 81-85 (2007)
Shubhakar, A., Kozak, R.P., Reiding, K.R., Royle, L., Spencer, D.I.R., Fernandes, D.L., Wuhrer, M. Automated High-Throughput Permethylation for Glycosylation Analysis of Biologics using MALDI- TOF-MS Analytical Chemistry. acs.analchem. 6b01639 (2016).
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