Vitronectin (SRP3186) is used to coat culture vessels for cells attachment. Below is a suggested coating protocol for a 96-well plate at a coating concentration of 0.5 µg/cm2. Note that the optimal concentration will vary with each cell line. For culture vessels other than a 96-well plate, scale this protocol accordingly. To calculate the working concentration of vitronectin, use Table 1, or the following equation:
Working concentration calculation Example:
To coat a 96-well plate (surface area is 0.3 cm2/well) at a coating concentration of 0.5 µg/cm2, prepare 9.6-10 mL of diluted vitronectin solution (0.1 mL/well × 96 wells, Table 1) at the following concentration:
The vitronectin working concentration in this example is 1.5 µg/mL. Adding 0.1 mL per well of this working solution will yield a coating concentration of 0.5 µg/cm2.
The biological activity of human, recombinant Vitronectin (VTN) expressed in HEK 293 cells (SRP3186) was measured in a cell attachment assay using CHO cells. The EC50 is defined as the effective concentration that elicits a 50% increase in cell attachment in a cell-based bioassay.
A. Preparation of vitronectin
B. Coating the plate