Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL)
Sensitivity
8 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
Specificity
Natural and recombinant human IFN-gamma
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Human IFN-gamma Immunoassay is a 4.5 hour solid phase ELISA designed to measure IFN-gamma levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IFN-gamma and antibodies raised against the recombinant factor. Results obtained for naturally occurring human IFN-gamma samples showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IFN-gamma.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Recovery
The recovery of IFN-gamma spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|
| Cell Culture Media (n=4) | 102 | 93-109 |
| EDTA Plasma (n=5) | 98 | 88-111 |
| Serum (n=5) | 102 | 91-118 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IFN-gamma in various matrices and diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
StorageStore the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IFN-gamma
IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.
Refer to the product for complete assay procedure.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
- Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample - Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate - Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
200 µL Substrate Solution
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
微博|
手机
