Human B-Raf Proto Oncogene Serine/Threonine Protein Kinase ELISA Kit

Manufactured by Antibodies.com, this is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) intended for the in vitro quantitative determination of human B-Raf Proto Oncogene Serine/Threonine Protein Kinase (BRAF) in tissue homogenates, cell lysates, or other biological fluids. It is listed under the Assays and Kits product category and is reactive with human samples.

The assay uses a sandwich enzyme immunoassay format on a pre-coated 96-well microplate arranged as 12 x 8 well strips. Standards and test samples are added to the wells, where BRAF present in each sample is captured by the immobilised antibody. After washing, a Biotinylated Anti-BRAF Antibody is added, followed by an HRP-Avidin conjugate. A TMB substrate solution is then used to visualise the enzymatic reaction, producing a blue-coloured product that changes to yellow after the addition of acidic stop solution. The intensity of yellow colour is proportional to the amount of BRAF captured in each well. The concentration of BRAF is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

Manufacturer-stated performance figures include an assay range of 0.312–20 ng/ml and a sensitivity of <0.124 ng/ml, with a total assay time of 4 h 30 min. Detection is colourimetric, read on a colorimetric microplate reader. Reported recovery values (n=5) were 80–102% for serum, 81–100% for EDTA plasma, and 80–89% for heparin plasma. Dilution linearity data across 1:2, 1:4, 1:8, and 1:16 dilutions were also reported for all three sample types. Kit components should be stored at +4°C or −20°C as instructed, and the kit should not be used past its expiration date.