Porcine Arachidonate 12-lipoxygenase, 12S-type (ALOX12) ELISA Kit

The Assay Genie Porcine ALOX12 Sandwich ELISA Kit enables quantitative measurement of arachidonate 12-lipoxygenase, 12S-type in porcine serum and plasma using a colorimetric detection method. ALOX12 catalyzes the oxygenation of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid, a key mediator in inflammatory and platelet signaling pathways, supporting cardiovascular, metabolic, and immune research in swine models. Each kit contains a pre-coated 96-well microplate, standard curve reagents, and assay buffers for full protocol execution. Samples should be stored refrigerated at 2-8°C and protected from repeated freeze-thaw cycles to preserve analyte integrity. The assay is intended for in vitro research use only and provides validated performance specifically for porcine sample matrices, with a standard curve range and sensitivity calibrated for endogenous lipoxygenase quantification.

Assay Genie's ALOX12 immunoassay leverages a pair of high-affinity monoclonal capture and detection antibodies directed against conserved epitopes of porcine platelet-type 12-lipoxygenase, minimizing cross-reactivity with ALOX5 and ALOX15 isoforms. The complete workflow typically completes within 2.5 to 3 hours at room temperature, with a recommended sample volume of 50 to 100 µL per well. Absorbance is read at 450 nm using a 620 nm or 630 nm reference wavelength to correct for plate well optical irregularities. Each lot is calibrated against a recombinant porcine ALOX12 reference standard, with an analytical sensitivity generally below 0.1 ng/mL and intra-assay and inter-assay coefficients of variation under 10% and 12%, respectively.

Because ALOX12-generated 12(S)-HETE and 12-HPETE serve as upstream regulators of platelet aggregation, vascular smooth muscle proliferation, and neutrophil chemotaxis, this Sandwich ELISA is widely used in translational studies of thrombosis, atherosclerosis, non-alcoholic fatty liver disease, and obesity-related metabolic dysfunction in swine. Reconstituted standards and biotinylated detection antibody should be stored at -20 degrees C in single-use aliquots to prevent performance drift, while the pre-coated 96-well microplate remains stable at 2-8 degrees C in its sealed foil pouch with desiccant until the printed expiration date. Researchers preparing serum should avoid extended room temperature incubation and process samples within 2 hours of collection or freeze at -80 degrees C for longer-term archiving, since repeated freeze-thaw cycles degrade the lipoxygenase epitope recognized by the capture antibody.