The HTRF human IL6 kit is designed for the quantification of IL6 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 and disposal requirements under European REACH regulations (EC 1907/2006).
Feature | Specification |
---|---|
Application | Protein Quantification |
Sample Volume | 16 µL |
The HTRF human IL6 kit is designed for the quantification of IL6 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 and disposal requirements under European REACH regulations (EC 1907/2006).
IL6 is a pro-inflammatory cytokine involved in acute-phase reaction, inflammation, and cancer progression. Secreted by T cells, macrophages, and fibroblasts, it induces B and T cell proliferation. It is being studied in a wide variety of research areas including diabetes, Alzheimer's disease, depression, and several cancers. Along with TGF beta, IL6 is main promoter of T cell differentiation into TH17, a new component of immuno-oncology.
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.
Application |
Protein Quantification
|
---|---|
Brand |
HTRF
|
Detection Modality |
HTRF
|
Product Group |
Kit
|
Sample Volume |
16 µL
|
Shipping Conditions |
Shipped in Dry Ice
|
Target Class |
Cytokines
|
Target Species |
Human
|
Technology |
TR-FRET
|
Therapeutic Area |
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
|
Unit Size |
10,000 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.
Sample size | 16 µL |
---|---|
Final assay volume | 20 µL |
Kit components | Lyophilized standard, frozen detection antibodies, buffers &protocol. |
LOD &LOQ (in Diluent) | 8 pg/mL &34 pg/mL |
Range | 34 7,500 pg/mL |
Time to result | 2h at RT |
Calibration | NIBSC (89/548) value (IU/mL) = 0,1 x HTRF hIL6 value (pg/mL) |
Species | Human only |
Intra-assay (n=24)
Sample | Mean [IL6] (pg/mL) | CV |
---|---|---|
1 | 384 | 5% |
2 | 2522 | 3% |
3 | 8594 | 3% |
Mean CV | 4% |
Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.
Inter-assay (n=4)
Sample | [IL6] (pg/mL) | Mean (delta R) | CV |
---|---|---|---|
1 | 116.5 | 400 | 13% |
2 | 268 | 1805 | 10% |
3 | 3261 | 7834 | 2% |
Mean CV | 8% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
Sample | Dilution factor | [IL6] expected (pg/mL) | [IL6] 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 | - | 354 | - |
4.3 | 83 | 82 | 101% | |
7.4 | 45 | 48 | 94% | |
Mean | 97% |
The excellent % of recovery obtained from these experiments show the good linearity of the assay.
Sample | [IL6] added (pg/mL) | [IL6] expected (pg/mL) | [IL6] detected (pg/mL) | Recovery |
---|---|---|---|---|
1 | 3250 | 4098 | 4103 | 100% |
Recombinant cytokine was added to a serum sample, and the response obtained from a standard curve was compared to the calculated expected values. The 100% of recovery observed validates the sample matrix used for this assay.
THP1 cells plated at 100 kcells/well were incubated for 18 h with increasing concentrations of JTE 607 in presence of 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 IL6 Assay.
PBMC were plated at 50, 100 and 400 kcells/well and stimulated for 18 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 IL6 Assay.
Are you looking for resources, click on the resource type to explore further.
Discover the cutting edge of neuroinflammation assay research
This literature review illustrates the possibilities and versatility...
We are here to answer your questions.