EasyPro Series ELISA Kits
EasyPro Human Adrenocorticotropic Hormone(ACTH)ELISA Kit
Product No
FY-EH1070SSample Type
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μLAssay Time
45minPurchase details
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DataSheet 
Product Details
Product Name in English:
EasyPro Human Adrenocorticotropic Hormone(ACTH)ELISA Kit
Species:
Human
Alternative Name:
Corticotropin, Adrenocorticotropin
Uniport ID:
P63267
Assay Type:
Competitive
Standard:
1000pg/mL
Sensitivity:
9.38 pg/mL
Detection Range:
15.63-1000pg/mL
Assay Time:
45min
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μL
Research:
Signal transduction;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ACTH Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for ACTH are added to the micro ELISA plate wells. ACTH in samples (or standards) competes with a fixed amount of ACTH on the solid phase supporter for sites on the HRP linked detection antibody specific to ACTH. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The concentration of ACTH in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration | OD1 | OD2 | Average OD | Corrected |
|---|---|---|---|---|
| 1000 | 0.258 | 0.258 | 0.258 | |
| 500 | 0.358 | 0.362 | 0.36 | |
| 250 | 0.552 | 0.56 | 0.556 | |
| 125 | 0.813 | 0.821 | 0.817 | |
| 62.5 | 1.108 | 1.127 | 1.118 | |
| 31.25 | 1.503 | 1.53 | 1.517 | |
| 15.63 | 1.817 | 1.825 | 1.821 | |
| 0 | 2.481 | 2.501 | 2.491 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
| Item | Intra-assay Precision | Intra-assay Precision |
|---|---|---|
| Sample | 3 | 3 |
| Replicate | 9 | 18 |
| CV(%) | 5 | 8 |
Rate of recovery
The recovery of 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) | 87-97 | 90 |
| EDTA plasma (n=8) | 81-95 | 88 |
| Cell culture media (n=8) | 83-95 | 89 |
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-92 | 86-99 | 86-97 |
| Average (%) | 86 | 87 | 88 | |
| 1:4 | Range (%) | 80-96 | 93-107 | 88-95 |
| Average (%) | 80 | 96 | 88 | |
| 1:8 | Range (%) | 89-100 | 85-94 | 92-101 |
| Average (%) | 98 | 90 | 100 | |
| 1:16 | Range (%) | 85-96 | 87-101 | 87-98 |
| Average (%) | 90 | 99 | 88 |
Assay Procedures
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