Cell source Stimulation Secretion stop Fixation Permeabilisation Staining Protocol



We define two ways of stimulation; the polyclonal and (antigen) specific stimulation. Stimulating cells polyclonal means that approximately every cell in the population is activated. Or at least can be activated. The second way of stimulation is meant to activate a part of the population you work with. 

Polyclonal stimulation

There are many ways of stimulating cells polyclonal. It is also the most often used way of stimulation to do intracellular staining. You can think of the already mentioned PMA/ionomycin stimulation which activates almost any cell to produce any protein it can. This is also the most unphysiologic way of stimulation. You can also think of CD3/CD28 or lectin (PHA or ConA) stimulation of T cells or LPS stimulation of monocytes or keratinocytes.

Specific, monoclonal stimulation

This is the most physiologic way of stimulation. And you can think of the percentage antigen specific T or B lymphocytes in your PBMC. That way you got a screening method for changes due to immunization protocols i.e.

Stimulating PBMC with PMA/ionomycin means every cell can be activated (B and T lymphocytes, NK cells and monocytes). Stimulating the same population with CD3/CD28 means only the T cells will be activated. In the second example only part of the population is activated but it still is an polyclonal activation.


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Page was last updated: 05-03-2005