Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL)
Sensitivity
7.5 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human IL-8
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
QC23, Quantikine Immunoassay Control Group 8 -
Please InquireProduct Summary
The Quantikine Human IL-8 Immunoassay is a 3.5 hour solid phase ELISA designed to measure human IL-8 in cell culture supernates, serum, and plasma. It is based on antibodies raised against the 72 aa variant of human IL-8 derived from E. coli. It is calibrated with the same recombinant factor. This immunoassay accurately quantitates recombinant human IL-8. Measurement of natural human IL-8 or the 77 aa variant of human IL-8 gave results parallel to the standard curves obtained using the E. coli-expressed Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-8.
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) hree 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 | 168 | 526 | 1093 | 196 | 581 | 1160 |
| Standard Deviation | 9.4 | 28.6 | 70.7 | 14.5 | 56.6 | 70.2 |
| CV% | 5.6 | 5.4 | 6.5 | 7.4 | 9.7 | 6.1 |
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision |
|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 115 | 386 | 802 | 132 | 410 | 817 |
| Standard Deviation | 5.3 | 17 | 37.9 | 10.7 | 28 | 42.4 |
| CV% | 4.6 | 4.4 | 4.7 | 8.1 | 6.8 | 5.2 |
Recovery
The recovery of IL-8 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|
| Cell Culture Media (n=5) | 98 | 85-114 |
| Citrate Plasma (n=5) | 105 | 95-114 |
| EDTA Plasma (n=5) | 103 | 97-111 |
| Heparin Plasma (n=5) | 102 | 92-107 |
| Serum (n=5) | 98 | 88-106 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of IL-8 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: CXCL8/IL-8
CXCL8/Interleukin-8 is a chemokine that is upregulated at sites of inflammation where it promotes neutrophil infiltration and activation. CXCL8 can form homodimers and heterodimers with CXCL4/PF4. Its bioactivity is regulated by proteolytic truncations, citrullination, and the decoy receptor DARC. CXCL8 signals through CXCR1/IL-8 RA, which is also used by CXCL6, and through CXCR2/IL-8 RB, which is used by multiple CXC chemokines. CXCL8 also binds to Serpin A1/alpha-1 Antitrypsin which prevents CXCL8 interaction with CXCR1.
Entrez Gene IDs
3576 (Human); 396880 (Porcine); 403850 (Canine); 493836 (Feline);
Alternate Names
3-10C; AMCF-I; C-X-C motif chemokine 8; CXCL8SCYB8; Emoctakin; GCP1; GCP-1TSG-1; IL8; interleukin 8; K60; LAI; LECT; MDNCF; MDNCFb-ENAP; member 8; MONAPGCP1; NAP1; NAP-1NAP1; NCF; Neutrophil-activating protein 1; Protein 3-10C; T cell chemotactic factor; T-cell chemotactic factor; TCF; TSG1;
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.
50 μL Standard, Control, or Sample - Add 50 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.
100 μL Conjugate - Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
- 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.
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