Description
| Product SKU: | SBRS0147 |
| Size: | 96T |
| Application: | This kit detects 420aa Nucleobindin-2 protein. Cross Reactivity: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Angiotensin II, NPY and APC. |
| Uniprot: | Q9JI85 |
| Gene Names: | NEFA / NUCB2 |
| Synonyms: | Nucleobindin-2 (DNA-binding protein NEFA) (Prepronesfatin) [Cleaved into: Nesfatin-1] |
| Target Species: | Human, Mouse, Rat |
| Compatible Sample Types: | Cell Culture Supernatants, Serum |
| Design Principle: | Competition-based |
| Method of Detection: | Colorimetric |
| Quantitative/Semi-Quantitative: | Quantitative |
| Range: | 0.1-1,000 ng/ml |
| Sensitivity: | 147 pg/ml or 3.09 pM |
| Recommended Dilution: | Human: 4X, Mouse: 4X, Rat: 2X |
| Storage/Stability: | Standard, biotinylated peptide, and positive control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened microplate wells and antibody (Item N) may be stored for up to 1 month at 2 to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge. |
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- HRP-Streptavidin
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- Technical Manual
Other materials and equipment required:
The Assay Genie Rat Nucleobindin-2 PharmaGenie ELISA Kit (SBRS0147) will require other equipment and materials to carry out the assay. Please see list below for further details.
- Distilled or deionized water
- Precision pipettes to deliver 2 ul to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
- Prepare all reagents, samples and standards as instructed.
- Add 100 ul detection antibody to each well.
- Incubate 1.5 h at RT or O/N at 4°C.
- Add 100 ul standard or sample to each well.
- Incubate 2.5 h at RT.
- Add 100 ul prepared streptavidin solution.
- Incubate 45 min at RT.
- Add 100 ul TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 ul Stop Solution to each well.
- Read plate at 450 nm immediately.
Nucleobindin-2 has roles in calcium level maintenance, eating regulation in the hypothalamus, and release of tumor necrosis factor from vascular endothelial cells. Nucleobindin-2 binds calcium and has EF-folding domains. Nucleobindin-2 is further cleaved into a 82 amino acids peptide, Nesfatin, which plays important roles regulating food intake, energy homeostasis, and blood pressure.
