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Human ACTH(Adrencocorticotropic Hormone) ELISA Ki

Human ACTH(Adrencocorticotropic Hormone) ELISA Ki
Human ACTH(Adrencocorticotropic Hormone) ELISA Ki
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Product Code : E-EL-H0137
Brand Name : Human ACTH
Product Description

Intended use

This ELISA kit can be applied to the in vitro quantitative determination of Human ACTH concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with Human ACTH. During the reaction, Human ACTH in the sample or standard competes with a fixed amount of Human ACTH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human ACTH. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Human ACTH in the samples is then determined by comparing the OD of the samples to the standard curve.

Assay type Competitive
Format 96T
Assay time 2.5h
Reactivity Human
Detection Method Colormetric
Detection Range 15.63—1000 pg/mL
Sensitivity 9.38 pg/mL
Sample Volume 50μL
Sample Type Serum, plasma and other biological fluids

Specificity

This kit recognizes Human ACTH in samples. No significant cross-reactivity or interference was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human ACTH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human ACTH 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
(pg/mL)
45.70 130.90 461.60 45.10 124.70 448.60
Standard
deviation
3.20 6.80 14.30 2.30 6.40 19.70
C V (%) 7.00 5.19 3.10 5.10 5.13 4.39

Recovery

The recovery of Human ACTH  spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Range (%) Average Recovery (%)
Serum (n=5) 91-104 98
EDTA plasma (n=5) 85-100 91
Cell culture media (n=5) 86-98 93

 

Kit components & Storage

The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions since the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 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 4℃(shading light)
Stop Solution 1 vial, 10 mL 4℃
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use in measuring instead of directly pouring.


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