Sheep Estrogen ELISA Kit
- SKU:
- SHFI00063
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.375pg/ml
- Range:
- 15.625-1000pg/ml
- ELISA Type:
- Competitive ELISA, Coated with Antigen
- Synonyms:
- ESRRG, Estrogen-related receptor gamma, ERR3, NR3B3, ERR gamma-2, ERRG2, ERRgamma, Estrogen receptor-related protein 3, NR3B3, Nuclear receptor subfamily 3 group B member 3, KIAA0832
- Reactivity:
- Sheep
Description
Kit Protocol
1. | When diluting samples and reagents, they must be mixed completely and evenly. Before adding TMB into wells, equilibrate TMB Substrate for 30 minutes at 37°C. It is recommended to plot a standard curve for each test. |
2. | Set standard, test samples, control (blank) wells on the pre-coated plate respectively, and then, records their positions. |
3. | Add 50ul of Standard, Blank, or Sample per well. The blank well is added with Sample/Standard Dilution Buffer. Immediately add 50ul Biotin-labeled Antibody Working Solution into each well. Cover with the Plate sealer we provided. Gently tap the plate to ensure thorough mixing. Incubate for 45 minutes at 37°C. |
4. | Remove the cover, and wash plate 3 times with Wash Buffer, and let the wash buffer stay in the wells for 1 minute each time. After the last wash, remove any remaining Wash Buffer by aspirating or decanting |
5. | Add 100ul SABC Working Solution into each well. Cover it with a new Plate sealer. |
6. | Remove the cover and wash plate 5 times with Wash Buffer, and let the wash buffer stay in the wells for 1-2 minutes each time. |
7. | Add 90ul TMB Substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 |
8. | Add 50ul Stop Solution into each well. The color will turn yellow immediately. The adding order of Stop Solution should be as the same as the TMB Substrate Solution |
9. | Read the O.D. absorbance at 450nm in Microplate Reader immediately after adding the stop solution. |
Regarding calculation, the standard curve can be plotted as the O.D.450 of each standard solution (Y) vs, the respective concentration of the standard solution (X). The target concentration of the samples can be interpolated from the standard curve. It is recommended to use some professional software to do this calculation, such as Curve Expert 1.3 or 1.4.
Note: If the samples measured were diluted, multiply the dilution factor to the concentrations from interpolation to obtain the concentration before dilution.
Kit Components
Item | Size | Storage |
ELISA Microplate(Dismountable) | 8x12 | 2-8°C/-20°C |
Lyophilized Standard | 2 vial | 2-8°C/-20°C |
Sample Dilution Buffer | 20ml | 2-8°C |
Biotin-labeled Antibody(Concentrated) | 60ul | 2-8°C(Avoid Direct Light) |
Antibody Dilution Buffer | 10ml | 2-8°C |
HRP-Streptavidin Conjugate(SABC) | 120ul | 2-8°C(Avoid Direct Light) |
SABC Dilution Buffer | 10ml | 2-8°C |
TMB Substrate | 10ml | 2-8°C(Avoid Direct Light) |
Text | 10ml | 2-8°C |
Wash Buffer(25X) | 30ml | 2-8°C |
Plate Sealers | 5 |
Other Materials and Equipement
The ELISA Genie Sheep Estrogen ELISA Kit will require other equipment and materials to carry out the assay. Please see list below for further details.
1. Microplate reader (wavelength:450nm)
2. 37°C incubator
3. Automated plate washer
4. Precision single and multi-channel pipette and disposable tips
5. Clean tubes and Eppendorf tubes
6. Deionized or distilled water
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.
Information | Description |
Serum | Place whole blood sample at room temperature for 2 hours or put it at 2-8°C overnight and centrifugation for 20 minutes at approximately 1000×g, Collect the supernatant and carry out the assay immediately. Blood collection tubes should be disposable, non-pyrogenic, and non-endotoxin. |