Hormone & Small Molecule ELISA Kits
3-NT / 3-Nitrotyrosine ELISA Kit
- SKU:
- UNFI0036
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Sensitivity:
- 0.938ng/ml
- Range:
- 1.563-100ng/ml
- ELISA Type:
- Competitive
- Synonyms:
- 3-NT, 3-Nitrotyrosine
- Reactivity:
- Universal
Description
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
| 3-NT, 3-Nitrotyrosine | |
| Product Name: | 3-NT (3-Nitrotyrosine) ELISA Kit |
| Product Code: | UNFI0036 |
| Size: | 96 Assays |
| Target: | 3-NT |
| Alias: | 3-NT, 3-Nitrotyrosine |
| Reactivity: | Universal |
| Detection Method: | Competitive ELISA, Coated with Antibody |
| Sensitivity: | 0.938ng/ml |
| Range: | 1.563-100ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
Additional Information
| New Column | ||||||||||||||||||||||
| Recovery | Matrices listed below were spiked with certain level of 3-NT and the recovery rates were calculated by comparing the measured value to the expected amount of 3-NT in samples.
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 3-NT and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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| Intra-Assay: | CV <8% | |||||||||||||||||||||
| Inter-Assay: | CV <10% |
Protocol
*Note: Protocols are specific to each batch/lot.
For the exact instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blank well is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody working solution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thorough mixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA plate well, avoid touching plate walls and foaming). |
| 3. | Wash: Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 350µL) using a squirt bottle, multi-channel pipette, manifold dispenser or automated washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC working solution to each well. Cover with a new Plate sealer. Incubate for 30 minutes at 37°C. |
| 5. | Wash: Repeat the aspiration/wash process for five times. |
| 6. | TMB Substrate: Add 90µL of TMB Substrate to each well. Cover with a new plate sealer. Incubate for about 10-20 minutes at 37°C. Protect from light. The reaction time can be shortened or extended according to the actual color change, but not more than 30 minutes. When apparent gradient appeared in standard wells, you can terminate the reaction. |
| 7. | Stop: Add 50µL of Stop Solution to each well. Color turn to yellow immediately. The adding order of stop solution should be as the same as the substrate solution. |
| 8. | OD Measurement: Determine the optical density (OD Value) of each well at once, using a microplate reader set to 450 nm. You should open the microplate reader ahead, preheat the instrument, and set the testing parameters. |
Sample Preparation
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum: | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum and assay promptly or aliquot and store the samples at-80°C. Avoid multiple freeze-thaw cycles. |
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