Human V-Ki-Ras2 Kirsten Sarcoma Viral Oncogene Homolog ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of human KRAS in serum, plasma, tissue homogenates or other biological fluids. The assay uses a pre-coated 96-well microplate arranged as 12 x 8 well strips. An antibody specific for KRAS is immobilised on the plate; standards and samples are added and KRAS is captured, followed by incubation with a Biotinylated Anti-KRAS Antibody and then HRP-Avidin conjugate. TMB substrate is used to visualise the reaction, producing a blue product that turns yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader and concentration calculated against the standard curve.

Sensitivity is reported as 0.113 ng/ml, with an assay range of 0.312-20 ng/ml. Total assay time is 4h 30m. Components supplied include a pre-coated 96-well microplate (12 x 8 well strips) stored at -20°C, lyophilised standard in 2 vials (-20°C), Detection Solution A 120 µl (-20°C), Detection Solution B 120 µl (-20°C), 30X Wash Buffer 20 ml (+4°C), Sample Dilution Buffer 45 ml (-20°C), TMB Substrate 9 ml (+4°C), Stop Solution 6 ml (+4°C) and 5 adhesive plate sealers.

Spike-and-recovery in serum (n=5) averaged 91% across a range of 80%-102%; EDTA plasma (n=5) averaged 90% across 81%-100%; heparin plasma (n=5) averaged 84% across 80%-89%. Dilution linearity from 1:2 to 1:16 dilutions was 87-91%, 87-107%, 74-101% and 92-97% for serum (n=5); 90-105%, 84-101%, 90-101% and 79-108% for EDTA plasma (n=5); and 84-95%, 92-105%, 82-105% and 89-91% for heparin plasma (n=5). Kit components should be stored at +4°C or -20°C as instructed and not used past the stated expiration date.