Rat dGHRL(Desacyl Ghrelin) ELISA Kit (RTES01112)
- SKU:
- RTES01112
Description
Rat dGHRL(Desacyl Ghrelin) ELISA Kit
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat dGHRL . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat dGHRL and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat dGHRL, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat dGHRL. The concentration of Rat dGHRL in samples can be calculated by comparing the OD of the samples to the standard curve.
| Assay type | Sandwich |
| Format | 96T |
| Assay time | 4.5h |
| Reactivity | Rat |
| Detection Method | Colormetric |
| Detection Range | 15.63-1000 pg/mL |
| Sensitivity | 9.38 pg/mL |
| Sample Volume Required Per Well | 100uL |
| Sample Type | Serum, plasma and other biological fluids |
Specificity
This kit recognizes Rat dGHRL in samples. No significant cross-reactivity or interference between Rat dGHRL and analogues was observed.
Typical 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 (pg/mL) | O.D | Average | Corrected |
|---|---|---|---|
| 1000 | 2.441 2.459 | 2.45 | 2.382 |
| 500 | 1.583 1.609 | 1.596 | 1.528 |
| 250 | 0.977 0.961 | 0.969 | 0.901 |
| 125 | 0.467 0.485 | 0.476 | 0.408 |
| 62.5 | 0.253 0.247 | 0.25 | 0.182 |
| 31.25 | 0.176 0.162 | 0.169 | 0.101 |
| 15.63 | 0.117 0.123 | 0.12 | 0.052 |
| 0 | 0.065 0.071 | 0.068 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat dGHRL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat dGHRL 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) | 45.91 | 148.92 | 444.03 | 45.97 | 159.90 | 430.88 |
| Standard deviation | 2.56 | 8.83 | 19.18 | 3.02 | 9.47 | 21.37 |
| C V (%) | 5.58 | 5.93 | 4.32 | 6.57 | 5.92 | 4.96 |
Recovery
The recovery of Rat dGHRL 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=5) | 94-107 | 101 |
| EDTA plasma (n=5) | 91-104 | 99 |
| Cell culture media (n=5) | 86-101 | 93 |
Linearity
Samples were spiked with high concentrations of Rat dGHRL 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 (%) | 87-100 | 93-109 | 96-111 |
| Average (%) | 93 | 99 | 103 | |
| 1:4 | Range (%) | 90-106 | 88-98 | 83-93 |
| Average (%) | 96 | 93 | 88 | |
| 1:8 | Range (%) | 86-99 | 85-97 | 83-94 |
| Average (%) | 92 | 90 | 89 | |
| 1:16 | Range (%) | 93-105 | 85-97 | 88-101 |
| Average (%) | 98 | 90 | 94 |
Kit Components & Storage
An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20'C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 uL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 uL | -20'C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4'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 | 4'C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4'C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
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