Cytochemical experiments on lectin-displaying glycans

Summary

Lectin is a non-immunogenic glycoprotein that binds specifically to sugars, e.g., lectin from kidney beans binds only to glucose and mannose, and lectin from wheat germ binds only to N-acetylglucose. Moreover, lectins can also bind to markers such as colloidal gold, horseradish peroxidase, etc., so they can be used as a simple and selective probe for the detection of sugar groups. However, due to the complexity of glycans and the relative nature of lectin specificity, lectins show relatively crude glycan cytochemical results. Lectin distribution can be altered during cell maturation, in tumor cells, or when cells undergo damage. Source: Experimental Techniques and Applications of Immunohistochemistry

Operation method

basic program

Materials and Instruments

Tissue Samples
Lectin-colloidal gold probe Osmium acid

Move

1. Lectin-colloidal gold probes are prepared or purchased.

2. Tissue is routinely fixed, free cells are broken up, and tissue sections are thinned. 3.

3. rinse 3 times. 4.

4. Incubate with 5-25 μg/ml of lectin-colloidal gold at 15-25℃ for 30-60 min. 5.

5. rinse and osmium fixation as usual.

Caveat

1. The binding of the lectin to the sugar is an affinity cytochemistry and can therefore be referred to immunocytochemical methods.

2. If the lectin is a biotinylated lectin, it may be shown by ABC or affinity gold labeling.

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