Fluorescent derivatization of a protease antigen to track antigen uptake and processing in human cell lines

BMC Immunology

Table 2 In Vitro hydrolysis of FITC-Subtilisin to < 10 K by Preiss Cell Extracts^a

Cell Fraction	pH 5.2	pH 5.2	pH 7.7	pH 7.7
Cell Extract	+	-	+	-
Initial RFU	100	100	100	100
< 10 K Fraction	44	0.46	27	19

^aPreiss cell extracts (50 μl) were adjusted to pH 5.2 with 0.1 M sodium acetate or to pH 7.7 with 0.1 M sodium bicarbonate in a total volume of 1 ml and incubated with 15 μg of PMSF inactivated FITC-subtilisin (22 RFU/μg protein) at 37°C for 4 hr. The contents of each tube were centrifuged on a Centriprep YM-10 concentrator and the percentage of fluorescent activity recovered in the low molecular weight fraction calculated.

ISSN: 1471-2172