Mouse diamine oxidase,DAO ELISA Kit

Instructions
Code CSB-E10090m
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Mouse diamine oxidase (DAO) ELISA Kit is an enzyme-linked immunosorbent assay that quantitatively measures the content of mouse DAO in many biological fluids, including serum, plasma, and tissue homogenates. The kit has excellent sensitivity, high specificity, good linearity, precision low than 10%, high recovery, good lot-to-lot consistency. Get access to more details from the product instructions.

This assay employs the Sandwich-ELISA technique, in which a sandwiched complex is formed by DAO in the samples or standards, pre-coated anti-human DAO, and Biotin-labeled DAO antibody. The Ag-Ab-Ag immune complex is labeled with HRP-avidin and then develops a color reaction after the addition of the TMB substrate. The intensity of the color is positively correlated to the amount of DAO in the sample.

DAO is the main enzyme for the gastrointestinal degradation of histamine and other biogenic amines. It primarily degrades excess histamine in the human body. Histamine that either occurs naturally or from the diet can help to regulate specific functions of your digestive, nervous, and immune systems. DAO deficiency may result in histamine intolerance (HIT), a condition caused by elevated histamine levels in the body. DAO nutritional supplement is frequently used to treat symptoms of HIT.

Target Name amiloride binding protein 1 (amine oxidase (copper-containing))
Alternative Names Aoc1 ELISA kit; Abp1Amiloride-sensitive amine oxidase [copper-containing] ELISA kit; DAO ELISA kit; Diamine oxidase ELISA kit; EC 1.4.3.22 ELISA kit; Amiloride-binding protein 1 ELISA kit; Amine oxidase copper domain-containing protein 1 ELISA kit; Histaminase ELISA kit
Abbreviation DAO
Uniprot No. Q8JZQ5
Species Mus musculus (Mouse)
Sample Types serum, plasma, tissue homogenates
Detection Range 15.6 mU/mL-1000 mU/mL
Sensitivity 3.9 mU/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of mouse DAO in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
SampleSerum(n=4)
1:1Average %84
Range %81-87
1:2Average %95
Range %91-99
1:4Average %91
Range %88-96
1:8Average %96
Range %93-99
Recovery
The recovery of mouse DAO spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 109104-112
EDTA plasma (n=4)8581-89
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
mU/mlOD1OD2AverageCorrected
10002.879 2.752 2.816 2.607
5002.301 2.216 2.259 2.050
2501.584 1.560 1.572 1.363
1250.945 0.932 0.939 0.730
62.50.655 0.672 0.664 0.455
31.20.532 0.542 0.537 0.328
15.60.412 0.407 0.410 0.201
00.211 0.206 0.209
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-mouse DAO antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled DAO antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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Target Background

Function
(From Uniprot)
Catalyzes the degradation of compounds such as putrescine, histamine, spermine, and spermidine, substances involved in allergic and immune responses, cell proliferation, tissue differentiation, tumor formation, and possibly apoptosis.
Subcellular Location Secreted, extracellular space
Protein Families Copper/topaquinone oxidase family
Database Links

STRING: 10090.ENSMUSP00000031835

UniGene: Mm.213898

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