NucleoCounter® Reagents
To determine the total number of mammalian cells in a sample, the use of reagent A100 and subsequently Reagent B is necessary.
The lysis Reagent A100, with a pH of about 1.25, is used to disrupt the plasma membranes of the mammalian cells, rendering the nuclei susceptible to staining with propidium iodide (PI). Secondly, the reagent contributes effectively to disaggregation of cell clusters.
The stabilizing Reagent B is used after Reagent A100 to raise the pH value, allowing PI to stain the DNA of the cells more efficiently.
It is recommended to use equal volumes of sample, Reagent A100 and Reagent B (e.g. 200 ml of each)