Affinity Analysis of Concanavalin A–Polysaccharides Interaction by Quartz Crystal Microbalance (QCM) / -Dissipation (QCM-D) (CAT#: STEM-MB-0762-CJ)

Introduction

Concanavalin A (ConA) is a lectin (carbohydrate-binding protein) originally extracted from the jack-bean. It is a member of the legume lectin family. It binds specifically to certain structures found in various sugars, glycoproteins, and glycolipids, mainly internal and nonreducing terminal α-D-mannosyl and α-D-glucosyl groups. ConA is a plant mitogen, and is known for its ability to stimulate mouse T-cell subsets giving rise to four functionally distinct T cell populations, including precursors to regulatory T cells; a subset of human suppressor T-cells is also sensitive to ConA.

Polysaccharides are major classes of biomolecules. They are long chains of carbohydrate molecules, composed of several smaller monosaccharides. These complex bio-macromolecules functions as an important source of energy in animal cell and form a structural component of a plant cell.

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Principle Applications Procedure Materials Notes Related Products

Principle

The basic principle of QCM is the piezoelectric effect of quartz crystal. Upon an alternating voltage applied to the two poles of the quartz crystal, the quartz crystal of QCM produces mechanical vibration or oscillation. This resonance is greatly sensitive to the thickness of the crystal/electrode system and the frequency of the acoustic frequency. Once the mass on the quartz crystal increases or decreases, the resonance will be disturbed and the resonance frequency of the quartz crystal will change. Thus, the mass variation on the quartz crystal electrode surface is transformed into the frequency variation of the output electric signal of the quartz crystal oscillation and is further transformed into the high-precision data by the computer.

Applications

Immunology/Inflammation; Biochemistry

Procedure

1. Sensor preparation, reduce the risk of damaging or contaminating the sensor surface.
2. Preparation of sample liquids.
3. Places the sensor into the flow cell and ensures perfect alignment.
4. Instant identification of specific sensors by unique ID number.
5. Using quarter turn bayonet type sealed flow cell.
6. Enter the experiment details in the information panel.
7. Display frequency and dissipation in real time.
8. Observe the bubble-free filling through the transparent window.
9. Electronic log file detects all steps in the measurement process.
10. Export and analyze the data.

Materials

• Sample: Proteins; Microbes; Nucleic Acids; Enzymes; Cells; Polymers; Small molecules; Antibody; Lipids; Virus-like particles (VLPs);
• Equipment: Quartz Crystal Microbalance (QCM) / Quartz Crystal Microbalance-Dissipation (QCM-D)
• Running buffer
• (Optional) PBS solution

Notes

1. QCM is an extremely sensitive mass balance that measures nanogram to microgram level changes in mass per unit area.
2. QCM detection limit is approximately 250 ng mL−1, which is similar to that of the existing enzyme-linked immunosorbent assay (ELISA) method.
3. Unlike traditional QCM, the QCM with an added D measures an additional parameter, namely the dissipation factor. so QCM-D has great advantages in providing binding kinetics information by virtue of its label-free breadth of applicability, low cost, and sensing unique mass distribution and viscoelastic behavior aspects of whole-cell responses in real time.

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