HTRF human IL8 kit is designed quantification of human IL8 release in cell supernatant.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Also called CXCL8, IL8 is a chemokine mainly produced by macrophages, T cells, and neutrophils. IL8 acts as a chemoattractant neutrophils, and promotes infiltration and activation at inflammation sites. IL8 is involved in several cancer types due to its ability to promote angiogenesis and cell proliferation, as well as to inhibit apoptosis.
Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.
Assay Points |
500
|
---|---|
Assay Target Type |
Kit
|
Assay Technology |
HTRF
|
Brand |
HTRF
|
Quantity |
1
|
Shipping Conditions |
Shipped in Dry Ice
|
Therapeutic Area |
Oncology & Inflammation
|
Unit Size |
500 Assay Points
|
Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Revvity low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.
The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.
Revvity also worked with Myassays.com to help you in your data analysis.
Sample size | 16 µL |
---|---|
Final assay volume | 20 µL |
Kit components | Lyophilized standard, frozen detection antibodies, buffers &protocol. |
LOD &LOQ (in Diluent) | 6 pg/mL &32 pg/mL |
Range | 32 4,000 pg/mL |
Time to result | 1h at RT |
Calibration | NIBSC (89/520) value (IU/mL) = 0,01 x HTRF hIL8 value (pg/mL) |
Species | Human only |
Intra-assay (n=24)
Sample | Mean [IL8] (pg/mL) | CV |
---|---|---|
1 | 58 | 9% |
2 | 314 | 4% |
3 | 2074 | 2% |
Mean CV | 5% |
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
Inter-assay (n=4)
Sample | [IL8] (pg/mL) | Mean (delta R) | CV |
---|---|---|---|
1 | 78 | 273 | 2% |
2 | 376 | 1504 | 12% |
3 | 1818 | 5990 | 5% |
Mean CV | 6.3% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
Sample | Dilution Factor | [IL8] expected (pg/mL) | [IL8] detected (pg/mL) | Recovery |
---|---|---|---|---|
1 | 1 | - | 2015 | - |
4.3 | 469 | 489 | 104% | |
7.4 | 271 | 292 | 108% | |
10.7 | 188 | 210 | 111% | |
Mean | - | 108% | ||
2 | 1 | 399 | - | |
4.3 | 93 | 85 | 91% | |
7.4 | 54 | 56 | 103% | |
Mean | 97% |
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
Sample | [IL8] added (pg/mL) | [IL8] expected (pg/mL) | [IL8] detected (pg/mL) | Recovery |
---|---|---|---|---|
1 | 5000 | 5179 | 5008 | 97% |
2 | 5000 | 6047 | 5682 | 94% |
The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery found validates the sample matrix used for this assay.
THP1 cells plated at 100 k cells/well were incubated with increasing concentrations of JTE 607 for 18h, then stimulated for 3 h with 2 µg/mL LPS. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL8 Assay.
PBMC plated at 50, 100, 200, and 400 k cells/well were stimulated for 3 h with increasing concentrations of LPS (0, 0.02, 0.2, 2 µg/mL). 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL8 Assay.
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