This assay recognizes recombinant and natural rat DAO

Rat diamine oxidase (DAO) ELISA Kit


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Background

Diamine oxidase (DAO) is an enzyme synthesized primarily in the gastrointestinal mucosal cells. Serum levels of DAO have been used as an indicator of the integrity and/or functional mass of the intestinal mucosa. The enzyme is also produced by the placenta and is elevated in newborn serum.

Feature and Advantages

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to DAO. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for DAO and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain DAO, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of DAO in the samples is then determined by comparing the O.D. of the samples to the standard curve.

< SPECIFICITY>
This assay recognizes recombinant and natural rat DAO. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
0.156 ng/ml-10 ng/ml
The standard curve concentrations used for the ELISA’s were
10 ng/ml, 5 ng/ml, 2.5 ng/ml, 1.25 ng/ml, 0.625 ng/ml, 0.312 ng/ml, 0.156 ng/ml.

<SENSITIVITY>
The minimum detectable dose of rat DAO is typically less than 0.039 ng/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Kit component

  Reagent Quantity
1. Assay plate 1
2. Standard 2
3. Sample Diluent 1 x 20 ml
4. Biotin-antibody Diluent 1 x 10 ml
5. HRP-avidin Diluent 1 x 10 ml
6. Biotin-antibodyt 1 x 120μl
7. HRP-avidin 1 x 120μl
8. Wash Buffer 1 x 20 ml (25×concentrate)
9. TMB Substrate 1 x 10 ml
10. Stop Solution 1 x 10 ml
Product List

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Species : 0.156 ng/ml - 10 ng/ml
Assay range : 100 µL (Cell Culture Supernates, Plasma,Serum)
Assay sample volume : 6-8 hr.+3 hr


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