Human Plexin B1 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, designed for the in vitro quantitative determination of human Plexin B1 in tissue homogenates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, in which an antibody specific for Plexin B1 is pre-coated onto a 96-well microplate. Standards and test samples are added to the wells and Plexin B1 is captured by the immobilised antibody. Following incubation and washing, a Biotinylated Anti-Plexin B1 Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of the acidic stop solution. The intensity of yellow is proportional to the amount of Plexin B1 captured in each well, and is read as optical density at 450nm in a microplate reader against a standard curve.

The kit has an assay range of 1.56–100 ng/ml and a sensitivity of 0.59 ng/ml, with a total assay time of approximately 4 hours 30 minutes. Validation data for serum (n=5), EDTA plasma (n=5) and heparin plasma (n=5) demonstrate recoveries of 80–102% (average 91%), 81–101% (average 91%) and 80–89% (average 84%) respectively. Linearity has been demonstrated across serial two-fold dilutions (1:2 to 1:16) in these matrices. Kit components are stored at +4°C or –20°C as indicated on the component labels. The product is intended for research use only and is not for diagnostic or therapeutic use.

Required materials not included with the kit include a microplate reader capable of measuring absorbance at 450nm ± 10nm, a multichannel pipette reservoir or automated microplate washer, software capable of generating logarithmic functions, multi- and single-channel pipettes with sterile tips, test tubes and Eppendorf tubes, deionised or distilled water, absorbent paper towels, and a 37°C incubator. The Stop Solution provided is an acid solution and the TMB Substrate is light sensitive; appropriate protective equipment such as gloves, goggles, masks and lab coats should be worn during use.