Question

You are working with a mixed population of cells that includes some cells expressing CD95 on their surfaces. Design an experimental protocol using Fluorescence-Activated Cell Sorting (FACS) to determine how many CD95-expressing cells are dividing at any given time.

Answer 1

Fluorescence-Activated Cell Sorting or FACS is a method that allows us to sort out different cell types on the basis of fluorescence emitted by cell membrane components bound compounds.

In the case of CD95 integral protein which is known for highly glycosylated, if use antibodies against cells with CD95 marker, will not identify glycans and can’t be targeted by antibodies as antibodies only recognize protein structures.

Therefore, two different proteins should be used in order to get the correct sorting of cells. One of these would bind to these glycosylations, and the other would be antibodies (with a fluorescent tag) against first.