Grade Anti-Human Inos Serum (Rabbit) (RIA) Antibody – Rabbit Polyclonal

Radioimmunoassay Procedure:
To 12 x 75 mm tubes are added 0.10 ml of either buffer. standard or unknown and 0.10 ml of antiserum after rehydrating the lyophilized powder to 20 ml with assay buffer. The assays are incubated overnight at 4 degree C before 0.10 ml of 125I-hiNOS [Tyr1136-1137-1153] which contains about 50.000 cpm is added or 0.10 ml of 125I-hiNOS which contains about 100.000 cpm is added. The assays are incubated for an additional 24 hr before bound tracer is separated from free tracer. There are two recommended procedures for the separation of bound from free tracer: (1) by precipitation with goat anti-rabbit IgG antiserum and carrier normal rabbit IgG followed by centrifugation and aspiration or (2) by binding to mag-beads coated with goat anti-rabbit IgG antiserum followed by collection of the mag-beads on a magnet and decanting the supernatant. If possible. all dilutions and additions should be made in the RIA buffer: 0.1 M NaCl. 50 mM sodium phosphate. pH 7.2. 10 mM Na2EDTA. 1.0% BSA. 0.1% Triton-X100. and 0.01% NaN3 which contains 200 KIU aprotinin/ml.
Standard range: 1.0 – 128 fmole hiNOS [1137-1153] per tube
Maximum sensitivity: 4 fmole hiNOS [1137-1153] per tube