Species:
Universal
Alternative Name:
ACT-B; PS1TP5BP1; Actin, cytoplasmic 1; Actin, cytoplasmic 1, N-terminally processed
Assay Type:
Competitive
unit:
ng/mL
Standard:
10 ng/mL
Sensitivity:
0.057 ng/mL
Detection Range:
0.16-10 ng/mL
Assay Time:
1.5H
Sample Type:
Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Signal transduction;Developmental science;Bone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ACTb. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to ACTb. 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 ACTb in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 10 | 0.266 | |
| 5 | 0.366 | |
| 2.5 | 0.56 | |
| 1.25 | 0.821 | |
| 0.63 | 1.116 | |
| 0.31 | 1.511 | |
| 0.16 | 1.825 | |
| 0 | 2.489 |
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) | 0.2 | 0.42 | 2.81 | 0.2 | 0.31 | 3.5 |
| Standard deviation | 0.02 | 0.05 | 0.15 | 0.03 | 0.06 | 0.18 |
| C V (%) | 3.27 | 7.57 | 2.63 | 5.05 | 4.42 | 4.07 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 83-96 | 89 |
| EDTA plasma (n=8) | 92-107 | 99 |
| Cell culture media (n=8) | 88-102 | 95 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 95-102 | 87-97 | 89-98 |
| Average (%) | 96 | 88 | 93 | |
| 1:4 | Range (%) | 83-96 | 87-97 | 90-99 |
| Average (%) | 91 | 93 | 91 | |
| 1:8 | Range (%) | 97-103 | 86-92 | 95-106 |
| Average (%) | 101 | 92 | 101 | |
| 1:16 | Range (%) | 85-92 | 89-103 | 87-101 |
| Average (%) | 86 | 101 | 87 |