How to prepare working solution of Phytohaemagglutinin (PHA) obtained from sigma aldrich for carrying out cytogenetics procedure?
Leukocyte cultivation medium: I used 55,5 µl of fhytohemaglutinin HA 15 (1800x, 9 mg/ml, Sigma-Aldrich) for 100 ml of complete medium.
Dear Gundeep Brar
PHA-M must first be reconstituted in sterile redist. water (final concentration of 2-10 mg/ml). This solution is stable between -15 to -25 degrees Celsius for several months when stored in aliquots (or for 2 weeks at +2 to +8 degrees). Further dilution is made with medium or PBS (phosphate buffer saline).
Optimum activity for stimulation of proliferation of lymphocytes is at 5-10 ug/ml.
As a working concentration used for stimulation of human peripheral blood lymphocytes, 2-10 ug/ml is recommended.
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