The cutinoglobulin A (ConA) agglutination assay can be used to: detect lectin receptors on the surface of cancer cells.

Operation method

Knife bean globulin A (ConA) agglutination assay

Principle

Lectins: Lectins are a class of proteins that bind specific sugar groups. Each lectin molecule has more than two binding sites. Cancer and transformed cells require a lower concentration of lectin than normal cells, which may be related to the distribution of lectin receptors on the cell surface; normal cells are dispersed, while cancer cells are more concentrated and have increased binding.

Materials and Instruments

Cells
PBS ConA
Oscillator Glass Plate

Move

1. Cell suspension preparation

Suspensions containing 10 × ¹⁰⁴ cells per milliliter were made with phosphate buffer solution (PBS) at pH 7.4 solution;
2. ConA PBS

0.59 % ConA solution was prepared in PBS. 3;
3. Aggregation test

Add cell suspension into several concave glass plate concave, 0.1 ml per concave, and then add ConA Add 0.1 ml of PBS to each concave glass plate, and then add 0.1 ml of ConA to each concave glass plate, so that the final concentration of ConA is 100, 50, 25, 12.5 and zero (control) μg;
4. Place on a micro-oscillator and oscillate for 5-10 minutes, then observe the agglutination phenomenon.
5. Results

If the cell suspension becomes flocculent, agglutination has occurred; cell agglutination is a marker for testing the biological properties of cells. Agglutination is a test of the biological properties of cells.

Caveat

If the cell suspension becomes flocculent, agglutination has occurred.

Common Problems

The cell agglutination reaction is a marker for testing the biological traits of cells.

Source : Tissue Culture and Molecular Cytology Techniques (Beijing Publishing House).

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