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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human |
| Sensitivity | 2.24 ng/ml |
| Detection Range | 2.24 ng/ml - 600 ng/ml |
| Specificity | This ELISA antibody pair detects human ATc3. Other species not determined. |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum |
| Recommended Dilution (Serum/Plasma) | 20 fold |
| Accession Number | Q12968 |
| Gene Id | 4775 |
| Gene Symbols | NFATC3, NFAT4 |
| Protein Name / Synonyms | Nuclear factor of activated T-cells, cytoplasmic 3 (NF-ATc3) (NFATc3) (NFATx) (T-cell transcription factor NFAT4) (NF-AT4) |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Inflammation |
| Shipping Type | Blue ice |
| Storage | -20°C |
Product Features
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
Standard Curves
Spiking & Recovery Results
| Sample Type | Average % Recovery | Range (%) |
|---|---|---|
| Serum | 106.8 | 89-121 |
| Plasma | 98.64 | 81-124 |
| Cell culture media | 103.6 | 89-122 |
Linearity Results
| Sample Type | Serum | Plasma | Cell Culture Media | |
|---|---|---|---|---|
| 1:2 | Average % of Expected | 109.9 | 121.6 | 122.1 |
| Range (%) | 108-112 | 113-130 | 112-132 | |
| 1:4 | Average % of Expected | 112.2 | 124.7 | 116.6 |
| Range (%) | 110-114 | 123-127 | 109-124 | |
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Stop Solution
- Assay Diluent(s)
- Lyophilized Standard
- Biotinylated Detection Antibody
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 µl volumes
- Adjustable 1-25 µl pipettes for reagent preparation
- 100 µl and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
- Prepare all reagents, samples and standards as instructed in the manual.
- Add 100 µl of standard or sample to each well.
- Incubate 2.5 h at RT or O/N at 4°C.
- Add 100 µl of prepared biotin antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared Streptavidin solution to each well.
- Incubate 45 min at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Storage/Stability
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.