Immune cell populations have unique functions. In order to study them, immune cells first need to be purified. Complement depletion is a common purification strategy. This method is rapid, low cost and high yield.
Purification of cell populations is a common procedure in our daily experiments. Here, Creative Biolabs describes a method and basic protocol for the rapid and efficient depletion of cell populations using antigen-specific antibodies and complement depletion.
|Solutions and Reagents
|Antigen-specific antibodies, rabbit complement, culture medium
The complement system consists of a large number of plasma proteins. When the complement system is activated by antibodies, a protein hydrolysis cascade reaction is initiated. The complement cascade can cleave any cell population in an antigen-specific manner. Thus, we can achieve cells depletion by adding antibodies and complement to cell populations.
Collect cell samples. Then adjust the number of starting cell populations in the culture medium.
First, select antibodies that are effective in activating complement and test their activity. Then, add each antibody to the cell suspension at an appropriate predetermined concentration. Finally, mix the cells well by inverting them back and forth.
First calculate the amount of complement required. Thaw the complement and sterilize it. Then add the complement directly to the tube containing the cells and antibodies.
Adjust the volume of cells in the tube and incubate the cells. At the end of the incubation, centrifuge the cells and discard the supernatant. Then collect the cell precipitate by multiple washes. Finally, check the cell purity by comparing the cell population of interest before and after depletion.
Poor cell purity
Purification targeting specific cell populations is a common experimental need. Many strategies can be applied. Complement depletion can be performed quickly in any laboratory and does not require expensive equipment or reagents. If you would like to try this fast and effective protocol, you can order our products and follow the protocol.
For research use only. Not intended for any clinical use.