Human Urokinase-type Plasminogen Activator Receptor (PLAUR/uPAR) ELISA Kit

UPAR consists of three different domains of the Ly-6/uPAR/alpha-neurotoxin family. All three domains are necessary for high affinity binding of the primary ligand, urokinase. It has been possible to express uPAR recombinantly in CHO-cells and S2 cells. 4 out of 5 of the possible glycosylation sites are used in vivo giving the protein a molecular weight of 50-60 kDA. Recently the structure of uPAR was solved by X-ray crystallography in complex with an peptide antagonist and with its native ligand, urokinase.However the components of the plasminogen activation system have been found to be highly expressed in many malignant tumors, indicating that tumors are able to hijack the system, and use it in metastasis. Thus inhibitors of the various components of the plasminogen activation system has been sought as possible anticancer drugs.

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Human Urokinase-type Plasminogen Activator Receptor (PLAUR/uPAR) ELISA Kit has high sensitivity and excellent specificity for detection of Human PLAUR/uPAR. No significant cross-reactivity or interference between Human PLAUR/uPAR and analogues was observed.

This Human Urokinase-type Plasminogen Activator Receptor (PLAUR/uPAR) ELISA Kit employs a two-site sandwich ELISA to quantitate PLAUR/uPAR in samples. An antibody specific for PLAUR/uPAR has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPLAUR/uPAR present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PLAUR/uPAR is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PLAUR/uPAR bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.