Species:
Universal
Alternative Name:
Acetylcholine
Assay Type:
Competitive
unit:
pg/mL
Standard:
1000 pg/mL
Sensitivity:
5.31 pg/mL
Detection Range:
15.63-1000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Signal transduction;Infection immunity;Neuro science;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ACH. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to ACH. Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ACH in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 1000 | 0.279 | |
| 500 | 0.379 | |
| 250 | 0.573 | |
| 125 | 0.834 | |
| 62.5 | 1.129 | |
| 31.25 | 1.524 | |
| 15.63 | 1.838 | |
| 0 | 2.502 |
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.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean(ng/mL) | 19.69 | 42.02 | 280.9 | 20.49 | 31.45 | 349.65 |
| Standard deviation | 2.1 | 4.64 | 14.51 | 2.61 | 5.77 | 18.06 |
| C V (%) | 5.72 | 6.11 | 3.11 | 4.94 | 6.38 | 2.95 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-90 | 84 |
| EDTA plasma (n=8) | 85-97 | 91 |
| Cell culture media (n=8) | 80-93 | 86 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-96 | 86-101 | 92-105 |
| Average (%) | 87 | 101 | 104 | |
| 1:4 | Range (%) | 87-101 | 86-101 | 89-102 |
| Average (%) | 97 | 100 | 90 | |
| 1:8 | Range (%) | 89-102 | 86-101 | 92-101 |
| Average (%) | 90 | 92 | 95 | |
| 1:16 | Range (%) | 85-98 | 87-96 | 88-102 |
| Average (%) | 91 | 89 | 94 |