Mouse Leptin ELISA Assay
The Mouse Leptin ELISA Assay is For Research Use Only
Size: 1×96 wells
Dynamic Range: 10 ng/mL – 0.156 ng/ml
Incubation Time: 130 minutes
Sample Type: Plasma, Serum, Tissue Culture Fluids, Urine
Sample Size: 100 μL
The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the LPT present in samples reacts with the anti- LPT antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound Leptins by washing, the Detection Antibody, biotin conjugated anti-LPT is added and complexes are formed. Following a wash step, the horseradish peroxidase (HRP) conjugated Streptavidin is added and complexes are formed. After another washing step, the complexes are assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of LPT in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of leptin in the test sample. The quantity of LPT in the LPT sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
SPECIMEN COLLECTION AND HANDLING
Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.