In the Mouse and Rat Leptin BioAssay(TM) ELISA, standards, quality controls, and samples are incubated in microplate wells coated with anti-mouse leptin antibody. After 60 minutes incubation and washng, biotin-labeled polyclonal anti-mouse leptin antibody is added to the wells and incubated with immobilized antibody-leptin complex for 60 minutes. After another washing, Streptavidin-HRP conjugate is added. Ater 30 minutes incubation and the last washing step, the remaining conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by the addition of acidic solution and the absorbance of the resulting yellow product is measured spectrophotometrically at 450nm. The absorbance is proportional to the concentration of leptin. A standard curve is constructed by plotting the absorbance values versus the leptin concentrations of standards and concentrations of unknown samples are determined using this standard curve.
Kit Components:
L1670-30J1: Microtiter Plate 1x96 wells
L1670-30J2: Biotin-Labeled Antibody Concentrate (10x), 1x1.3ml
L1670-30J3: Streptavidin-HRP Conjugate, 1x13ml
L1670-30J4: Mouse Standard 1x1vial
L1670-30J5: Rat Standard 1x1vial
L1670-30J6: Quality Control, Mouse 2x1vial
L1670-30J7: Quality Control, Rat 2x1vial
L1670-30J8: Dilution Buffer, 2x13ml
L1670-30J9: Biotin-Antibody Diluent, 1x13ml
L1670-30J10: Wash Solution Concentrate (10x), 1x100ml
L1670-30J11: Substrate Solution (TMB), 1x13ml
L1670-30J12: Stop Solution, H2SO4, 1x13ml
Storage and Stability:
Store *L1670-30J4, L1670-30J5, L1670-30J6, L1670-30J7 powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 3 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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