Human Fc Fragment Of IgG Low Affinity IIIa Receptor ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of human CD16a, also referred to as the Fc Fragment Of IgG Low Affinity IIIa Receptor. The kit is intended for the quantitative measurement of human CD16a in serum, plasma, tissue homogenates, cell lysates or other biological fluids, and it uses a pre-coated 96-well microplate (12 x 8 well strips) onto which an antibody specific for CD16a has been immobilised. Standards and test samples are added to the wells, followed by a Biotinylated Anti-CD16a Antibody and an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of the acidic stop solution. Absorbance is read at 450 nm in a microplate reader and the concentration of CD16a is calculated by reference to the standard curve.

Reported performance includes an assay range of 78.1–5000 ng/ml and a sensitivity of <34 ng/ml. Mean recovery values given for the assay are 91% in serum (range 83–100%), 95% in EDTA plasma (range 91–99%) and 94% in heparin plasma (range 90–99%). Linearity has been demonstrated across sample dilution series of 1:2, 1:4, 1:8 and 1:16 in serum, EDTA plasma and heparin plasma. The total assay time is approximately 4 hours 30 minutes and detection is colourimetric, performed on a microplate reader.

Kit components include the pre-coated 96-well microplate, lyophilised standard, Detection Solution A, Detection Solution B, a 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Components are stored at +4°C or -20°C as instructed, and the kit is labelled for research use only. Sample collection guidance covers serum collected in serum separator tubes and centrifuged at 1000 x g for 15 minutes, plasma collected with EDTA or heparin as anticoagulant and centrifuged at 1000 x g for 15 minutes, cell culture supernatant clarified at 500 x g for 5 minutes, tissue homogenised in PBS (1 g tissue / 9 ml PBS) and centrifuged at 5000 x g for 5 minutes, and cell lysates centrifuged at 14,000 x g for 5 minutes, with aliquots stored at -20°C or -80°C and freeze/thaw cycles avoided.