Lectins overlays

Protein-Blot Analysis of Glycoproteins and Lectin Overlays... 

Meany, D. L., L. Hackler, H. Zhang, and D. Chan. 2011. Tyramide signal amplification for antibody-overlay lectin microarray A strategy to improve the sensitivity of targeted glycan profiling. /. Proteome Res. 10(3) 1425-31. 

The N-linked glycomoiety can be distinguished from O-linked structures in that the former is exclusive for mannose residues. Mannose is recognized by the lectin Con A. Moreover, the enzyme HRP is also an N-linked glycoprotein. Therefore, the simple procedure developed by Clegg (1982) is suggested for Con A overlays. 

As is illustrated in Sections A and B, HRP is a convenient detection system. Many lectins can be purchased directly conjugated to this and other enzymes (e.g., alkaline phosphatase), enabling a single step overlay. Alternatively, iodinated lectins are commonly used (see Fig. 2). In addition, a wide variety of biotin-lectin conjugates are also available and would be used much in the same manner as is described in Section A, i.e., a two-step procedure (see also Section D). 

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