Mouse Sjogren Syndrome Antigen B ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (ELISA) is designed for the in vitro quantitative determination of mouse SSB (Sjogren Syndrome Antigen B) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The assay employs the sandwich enzyme immunoassay technique: an antibody specific for SSB is pre-coated onto a 96-well microtiter plate, standards and test samples are added, and SSB present in each sample is bound by the immobilized antibody. After washing, a Biotinylated Anti-SSB Antibody is added, followed by HRP-Avidin conjugate. TMB substrate solution produces a blue colour that turns yellow upon addition of acidic stop solution; the density of yellow is proportional to the amount of SSB captured. The concentration of SSB is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The standard curve product range is 0.156–10 ng/ml with a sensitivity of <0.057 ng/ml. Total assay time is 4h 30m on a pre-coated microplate platform of 12 x 8 well strips, with colourimetric detection on a microplate reader. Components include a pre-coated 96-well microplate (12 x 8 well strips), lyophilized standard (2 vials), detection solution A (120 μl), detection solution B (120 μl), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml) and 5 adhesive plate sealers, with storage at +4°C or −20°C as instructed. Kit components should not be used past the stated expiration date.

Recovery performance has been characterised across sample matrices. Average recovery in serum (n=5) ranged from 80% to 102% (mean 91%); EDTA plasma (n=5) ranged from 81% to 100% (mean 90%); heparin plasma (n=5) ranged from 80% to 89% (mean 84%). Dilution linearity was assessed at 1:2, 1:4, 1:8 and 1:16. Serum samples (n=5) showed recoveries of 87–91%, 87–107%, 74–101% and 92–97%; EDTA plasma (n=5) gave 90–105%, 84–101%, 90–101% and 79–108%; heparin plasma (n=5) gave 84–95%, 92–105%, 82–105% and 89–91%.