Analysis Affinity of CD19 to Whole Blood B Cell by KinExA (CAT#: STEM-MB-0048-CJ)

Introduction

The incidence of hematological malignancies has been increasing over the past 30 years and accounts for about 9 percent of all cancers. Lymphomas are the most common type of hematologic malignancies. B-cell lymphomas are more common than T-cell lymphomas, accounting for approximately 85% of all non-Hodgkin lymphomas (NHL). CD19 is a transmembrane glycoprotein of the immunoglobulin (Ig) superfamily. It is expressed at different stages of B-cell development, beginning at the pre-B-cell stage and ending in early plasma cells. In addition, CD19 is widely expressed in B-cell malignancies, including CD20 positive [e.g., NHL and B chronic lymphocytic leukemia (B-ClL) ] and low or negative CD20 [e.g., B acute lymphocytic leukemia].

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Principle Applications Procedure Materials Notes Related Products

Principle

KinExA is a two-stage analysis system. In the first stage, a number of solutions are prepared, where one partner remains constant (constant binding partner, or CBP) and the other (titrant) is variable, usually serially diluted. As the titrant is added, the free CBP decreases and is analysed by a sophisticated and precise microfluorescence measurement device. The signal generated can be mathematically related to the affinity (KD) of the two molecules for each other, as well as the kinetic parameters of binding (kon) and dissociation (koff).

Applications

Oncology & Cancer; Immunology/Inflammation; Pharmacology

Procedure

1. preparation of the functionalized beads which will capture the analyte for measuremen.
2. preparation of a series of solutions consisting of a constant initial concentration of one component of the binary reaction and serial dilutions of the other reactant. The component that is kept constant is the constant binding partner (CBP) , and is the one which will be analyzed.
3. each reaction mixture is sampled and the fluorescence of free CBP bound to the capture beads is obtained for subsequent numerical analysis.

Materials

• Sample Type: Antibodies, Cells, Small Molecules, Proteins, DNA, Lipids, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)

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