Glycoprofile™ 標籤試劑盒

GlycoProfile™標籤試劑盒專為高效標籤N-連結、O-連結和糖基磷脂酰肌醇（GPI）錨定的聚糖而設計，您可選擇使用2-氨基苯甲酰胺（2-AB）或2-氨基苯甲酸（anthranilic acid; 2-AA） ^1,2。這些小型螢光團可增加聚糖的光譜吸收，改善 HPAE (高效能負離子交換色譜法)、^3-6 HPLC^4-7 和 ESI-MS (電漿譜質譜法) 方法的檢測。結合穩固，所產生的染料-聚糖結合物也很穩定，在標示後的分析過程中不會降解。建議使用 2-AB，以 HPAE、HPLC 及 ESI-MS 進行下游分析，但其靈敏度略低於 2-AA。2-AA 建議用於 SDS-PAGE，也適用於 HPAE、HPLC 和 ESI-MS。

圖 1. 用 2-AB 標示碳水化合物

Features and Benefits

• 每個試劑盒都是完整的，包含 2-AA 或 2-AB 染料、還原劑和溶劑。
• 足以製備多達 36 個混合聚糖樣品，包含 100 皮摩尔到 50 納摩尔的純化聚糖。
• 足以製備多達 36 個混合聚糖樣品，其中包含 100 皮摩尔到 50 納摩尔的純化聚糖。對於單一純聚糖，只要 5 皮摩尔就可以進行標籤，並在後續的 HPLC 分析中檢測出來。

  GlycoProfile™ Glycan Clean-Up Cartridges 建議搭配 GlycoProfile Labeling 套件使用，以進行標示後的清理。甘聚糖會吸附在膜上，而多餘的染料、單糖和二糖以及其他疏水性污染物則會通過膜。純化後的標示聚糖會被水洗離，並可進行後續的酵素消化或分析。

參考資料

1. Bigge J, Patel T, Bruce J, Goulding P, Charles S, Parekh R. 1995. Nonselective and Efficient Fluorescent Labeling of Glycans Using 2-Amino Benzamide and Anthranilic Acid. Analytical Biochemistry. 230(2):229-238. https://doi.org/10.1006/abio.1995.1468

2. Hardy M. Glycan labeling with the fluorophores 2-aminobenzamide and anthranilic acid, Techniques in Glycobiology, Dekker, Inc. NY (1997)..

3. Delaney J, Vouros P. 2001. Liquid chromatography ion trap mass spectrometric analysis of oligosaccharides using permethylated derivatives. Rapid Commun. Mass Spectrom.. 15(5):325-334. https://doi.org/10.1002/rcm.230

4. Anumula KR, Dhume ST. 1998. High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid. Glycobiology. 8(7):685-694. https://doi.org/10.1093/glycob/8.7.685

5. Townsend R, Lipniunas PH, Bigge C, Ventom A, Parekh R. 1996. Multimode High-Performance Liquid Chromatography of Fluorescently Labeled Oligosaccharides from Glycoproteins. Analytical Biochemistry. 239(2):200-207. https://doi.org/10.1006/abio.1996.0315

6. Royle L, Mattu TS, Hart E, Langridge JI, Merry AH, Murphy N, Harvey DJ, Dwek RA, Rudd PM. 2002. An Analytical and Structural Database Provides a Strategy for Sequencing O-Glycans from Microgram Quantities of Glycoproteins. Analytical Biochemistry. 304(1):70-90. https://doi.org/10.1006/abio.2002.5619

7. Guile GR, Rudd PM, Wing DR, Prime SB, Dwek RA. 1996. A Rapid High-Resolution High-Performance Liquid Chromatographic Method for Separating Glycan Mixtures and Analyzing Oligosaccharide Profiles. Analytical Biochemistry. 240(2):210-226. https://doi.org/10.1006/abio.1996.0351

8. Rudd PM, Mattu TS, Zitzmann N, Mehta A, Colominas C, Hart E, Opdenakker G, Dwek RA. 1999. Glycoproteins: Rapid Sequencing Technology for N-linked and GPI Anchor Glycans. Biotechnology and Genetic Engineering Reviews. 16(1):1-22. https://doi.org/10.1080/02648725.1999.10647969