Description
Intended Use:Â Ethos Biosciences Rat NGAL ELISA is an immunoassay for the quantitative determination of rat NGAL (rNGAL) in rat specimens. The assay is intended for research use; it is not intended for diagnostic purposes.
Description: Rat NGAL ELISA is an antigen capture antibody sandwich ELISA run in direct mode. It uses a recombinant rNGAL for standard and relies on two different antibodies raised against rNGAL. These antibodies show no cross-reactivity with either rat immunoglobulins or rat albumin.
The rNGAL ELISA Plate is supplied dry, but is ready to use. It is coated with anti-rNGAL Ab. In practice, the plate is first washed and then controls, standard dilutions and (diluted) samples are added.
During an overnight incubation, rNGAL is captured by the immobilized antibody. Washing the plate removes unreacted materials, and a second anti-rNGAL antibody conjugated with horseradish peroxidase (HRP) is added. Over a two-hour incubation period this antibody-conjugate binds with rNGAL bound to the plate.
Unreacted conjugate is removed by washing the plate, and bound conjugate is detected using a chromogenic substrate for HRP. After a short color development period, acid stopper is added, and the absorbance is measured using a plate reader.
The absorbance (determined at 450 nm) is directly proportional to the concentration of rNGAL in the standards and samples. Dose-Responses for standard dilutions of recombinant rNGAL, and those for serially diluted rat urine are colinear or parallel when plotted appropriately. This indicates that the standard and antibodies are appropriate for measuring rNGAL in rat samples.
The standard curve for this assay runs from 0.31-20 ng rNGAL/mL.
Kit Contents:
- 1 x 96-well Rat NGAL Assay Plate
- 2 x NHE-Casein Diluent
- 1 x Rat NGAL Standard
- 1 x Anti-Rat NGAL-HRP Conjugate
- 1 x TMB Color Developer
- 1 x Acid Color Stopper
- 1 x Instructions
All kit reagents are supplied in ready to use liquid form.
Note: Items 2 through 4 contain 0.05% Proclin 300 (active component Isothiazolinone) as preservative.
Color Stopper contains 2.0 N Sulfuric Acid. Refer to the MSDS online at Ethos Biosciences’ website for details (www.ethosbiosciences.com/msds).
Save and store all unused reagents and strips (in sealed pouch with included desiccant pack) for future use at 4ºC. Do not use this kit beyond its expiration date.
Standard Curve: