DLAP was prepared by pouring ten mL of sterile BioChrome
Vibrio medium (BCVM, Chromogenic medium for V.
The human histiocytic lymphoma cell line U937 was obtained from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany) and cultured in RPMI 1640 (Gibco by Life Technologies, Darmstadt, Germany), supplemented with 10% fetal calf serum (Biochrome
, Berlin, Germany), and 2mM L-glutamine (Gibco), at 37[degrees]C, 5% C[O.sub.2].
Finally, the plate was incubated with 100 [micro]l of PNPP for 30 min and applied to ELISA reader (Biochrome
, Cambridge, UK) at [OD.sub.450].
The lab is equipped with technologically most advanced dedicated amino acid analyzer, Biochrome
Cells were cultured at 2 x [10.sup.6] cells/mL in RPMI-1640 (Gibco BRL, Paisley, Scotland) medium with a supplement of 10% fetal calf serum (Biochrome
KG) and antibiotics (100IU/mL penicillin, 100 [micro]g/mL streptomycin).
Fetal calf serum (FCS) was obtained from Biochrome
KG (Berlin, Germany).
SST was digested by mixing the punched gel exposed to the test specimen with 60 [micro]L collagenase (Biochrome
AG, Berlin, Germany) at 500 [micro]g/mL in PBS followed by incubation at 35[degrees]C for 1.5 hours.
The DNA concentration ([DNA]) was measured in ng x [micro][l.sup.-1], and the quality measured through the ratio A260:A28o (Sambrook and Russell, 2001) using a spectrophotometer (WPA, Biowave, Biochrome