Sialo-Profiling Kit (α2-3, α2-6)

Sialylation plays a significant role in immune responses, cellular communication, pathogen recognition, cell survival, and cardiovascular disease. The differentiation of sialic acid linkages (α2-3 vs. α2-6) plays a crucial role in various diseases; for example, α2-3 sialic acid is involved in cancer progression, whereas α2-6 is involved in immune response regulation. The sialylation profiling kit includes four distinct Lectins, enabling detection and analysis of α2-3 and α2-6 sialic acid in cells and tissues. Sambucus nigra Lectin (SNA I) can recognize α2-6 sialic acid on sialylated glycans, whereas MAA II (Maackia amurensis Lectin) can recognize α2-3 on the surface. The other two Lectins (PNA & ECL) can recognize non-sialylated glycans by their unique sugar specificities. This kit provides other reagents and buffers needed for specific analyses, as well as technical support for experimental needs.

Kit Contents:

Specifications:

• Sambucus nigra Lectin (SNA/EBL I) - Recognizes α2-6 sialic acids
• Maackia amurensis Lectin (MAA/MAL I+II) - Recognizes α2-3 sialic acids
• Arachis hypogaea Lectin (PNA) - Recognizes T antigen, an O-glycan
• Erythrina cristagalli Lectin (ECL/ECA) - Recognizes terminal beta 1,4-Gal
• Streptavidin :: FITC conjugate for detection

Technical Information

• Recommended cell number to use is 2 – 5X106 cells per analysis.
• Recommended Lectin dilution is 5 – 30 µg / mL in PBS Buffer 1X, pH 7.4.
• The assay kit is recommended for 20 tests (each Lectin/component).
• Binding efficiency depends on target cell lines and the time of incubation.
• Avoid multiple freeze-thaw cycles.
• Concentrated amounts of Lectin conjugate can cause cytotoxicity.

Storage and Stability

Store frozen at -20°C in amber vials or covered with foil in appropriate aliquot sizes. Avoid freeze-thaw cycles. It can be stored at 2-8°C for short-term use. Clarify by centrifugation, if needed.

Application

1.Total/altered sialylation detection in cellular communication, pathogen recognition, cell survival, and cardiovascular disease (Live cells, Plasma, Serum, Tissues).
2.Lectins, enabling detection and analysis of (α2-3 and α2-6) sialic acid in cells and tissues.
3.Detection of T – antigen (O Glycans).
4.Flow/IHC-based detection.

References

1.Kaku H, Peumans WJ, Goldstein IJ. Isolation and characterization of a second lectin (SNA-II) present in elderberry (Sambucus nigra L.) bark. Arch Biochem Biophys. 1990 Mar;277(2):255-62.
2.Maveyraud L, Niwa H, Guillet V, Svergun DI, Konarev PV, Palmer RA, Peumans WJ, Rouge P, Van Damme EJ, Reynolds CD, Mourey L. Structural basis for sugar recognition, including the Tn carcinoma antigen, by the lectin SNA-II from Sambucus nigra. Proteins. 2009 Apr;75(1):89-103.
3.Bum Soo Kim, Kyung Taik Oh,1, Due Hyeon Cho, Yun Jung Kim, Wan Mo Koo, Kwang Hoon Kong, HaHyung Kim. A sialic acid-binding lectin from the legume Maackia fauriei: comparison with lectins from M. amurensis. Plant Science 167 (2004) 1315–1321.
4.Konami Y, Yamamoto K, Osawa T, Irimura T. Strong affinity of Maackia amurensis hemagglutinin (MAH) for sialic acid-containing Ser/Thr-linked carbohydrate chains of N-terminal octapeptides from human glycophorin A. FEBS Lett. 1994 Apr 11;342(3):334-8.
5.Jie Sun, Qing-li Yang*, jie Bi, Chu-shu Zhang, Li-na Yu and Feng Zhu. Purification and Identification of a Natural Lectin from the Seed of Peanut Arachis hypogaea. The Open Materials Science Journal, 2011, 5, 78-82
6.Erber, W. N., Asbahr, H., Meyer, B., Herrmann, R. P., & Davies, J. M. (1992). Peanut agglutinin (lectin from Arachis hypogaea) binding to hemopoietic cells: an immunophenotypic study using a biotin streptavidin technique. Pathology, 24(3), 173-176.