Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL), Urine (100 uL)
Sensitivity
10 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
Specificity
Natural and recombinant human MCP-1
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.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 - Please InquireProduct Summary
The Quantikine Human MCP-1 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure MCP-1 in cell culture supernates, serum, plasma, and urine. It contains E. coli-expressed recombinant human MCP-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human MCP-1. Results obtained using natural human MCP-1 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 MCP-1.
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.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
| Intra-Assay Precision | Inter-Assay Precision |
|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 76.7 | 364 | 1121 | 74.2 | 352 | 1076 |
| Standard Deviation | 6 | 17.1 | 54.4 | 5 | 20.5 | 49.4 |
| CV% | 7.8 | 4.7 | 4.9 | 6.7 | 5.8 | 4.6 |
Cell Culture Supernates, Urine
| Intra-Assay Precision | Inter-Assay Precision |
|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 78.2 | 360 | 1079 | 76 | 360 | 1086 |
| Standard Deviation | 3.3 | 17.7 | 63.5 | 4.5 | 17.2 | 48.5 |
| CV% | 4.2 | 4.9 | 5.9 | 5.9 | 4.8 | 4.5 |
Recovery
The recovery of MCP-1 spiked to three levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|
| Cell Culture Media (n=5) | 96 | 88-107 |
| Citrate Plasma (n=5) | 100 | 94-107 |
| EDTA Plasma (n=5) | 96 | 92-102 |
| Heparin Plasma (n=5) | 102 | 94-114 |
| Serum (n=5) | 103 | 92-113 |
| Urine (n=5) | 92 | 85-100 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of MCP-1 and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
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.
50 μL Assay Diluent for Serum & Plasma Samples Only
- For Serum & Plasma Samples Only: Add 50 μL of Assay Diluent to each well.
200 μL Standard, Control, or Sample - Add 200 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 twice for a total of 3 washes.
200 μL Conjugate - Add 200 μL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate & Urine Samples:Cover with a new plate sealer, and incubate at room temperature for 1 hour. - Aspirate and wash 3 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|
手機(jī)
