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Human DHEA-S(Dehydroepiandrosterone Sulfate) ELISA Kit

  • £41400
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Human DHEA-S(Dehydroepiandrosterone Sulfate) ELISA Kit

Detection Range 15.63-1000 ng/mL
Sensitivity 9.38 ng/mL



Product Details

Properties

Assay type Competitive-ELISA
Format 96T
Assay time 2.0h
Detection range 15.63-1000 ng/mL
Sensitivity 9.38 ng/mL
Sample type &Sample volume Serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes DHEA-S in samples. No significant cross-reactivity or interference between DHEA-S and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of DHEA-S concentrations in Serum, plasma and other biological fluids.

 

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with DHEA-S. During the reaction, DHEA-S in the sample or standard competes with a fixed amount of DHEA-S on the solid phase supporter for sites on the Biotinylated Detection Ab specific to DHEA-S. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of DHEA-S in tested samples can be calculated by comparing the OD of the samples to the standard curve.

 

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8℃
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

 

 

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level DHEA-S were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level DHEA-S were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 46.30 89.60 442.40 44.50 84.10 469.90
Standard deviation 3.20 4.70 14.60 2.40 5.00 22.60
CV (%) 6.91 5.25 3.30 5.39 5.95 4.81

 

 Recovery

The recovery of DHEA-S spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 88-101 93
EDTA plasma(n=8) 88-99 93
Cell culture media(n=8) 88-102 95

 

Target Information

Synonyms Androstenolone sulfate; Prasterone sulfate; Androst-5-en-3β-ol-17-one 3β-sulfate
Research Area  Signaling transduction

 

 Assay Procedures

elisa assay procedure 1

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results