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.

Is it possible to use saline solution instead PBS buffer to dilute PHA powder?

Thanks for your answers!