Complement C5a receptor assay for high throughput screening.
The complement C5a receptor on U937 cells, a human histiocytic lymphoma cell line, stimulated with dibutyryl-cAMP have been stabilized for at least 3 months at a dilute, ready to use concentration. [125I]-Bolton Hunter labeled C5a, (recombinant, human) has been prepared by reverse phase HPLC to 2200 Ci/mmol. Using a filtration binding assay the Kd from receptor saturation analysis is 10-40 pM and there are 50,000-100,000 receptor sites per cell. These reagents have permitted the development of a reliable, reproducible and convenient drug screening assay, in kit format, for compounds acting at the C5a receptor.
Harris
S R
SR
E.I.Du Pont de Nemours, Biotechnology Systems Division, Billerica, MA 01862.
Garlick
R K
RK
Miller
J J
JJ
Harney
H N
HN
Monroe
P J
PJ
D015936:Complement C5a / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical; D006801:Humans; D007457:Iodine Radioisotopes; D011869:Radioligand Assay; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement / Q000032:analysis*; D014407:Tumor Cells, Cultured
D007457:Iodine Radioisotopes; D044087:Receptor, Anaphylatoxin C5a; D011951:Receptors, Complement; D015936:Complement C5a