The HTRF mouse IL6 kit is designed for the quantification of mouse 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 & 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. When assaying, always follow recommended protocol and avoid highly haemolyzed samples.
Assay Points |
500
|
---|---|
Assay Target Type |
Kit
|
Assay Technology |
HTRF
|
Brand |
HTRF
|
Quantity |
1
|
Shipping Conditions |
Shipped in Dry Ice
|
Therapeutic Area |
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
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.
To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting,
Revvity also worked with Myassays.com to help you in your data analysis. You’ll be able to access a free online software to run your IL6 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 &19 pg/mL |
Range | 19 7,000 pg/mL |
Time to result | 1h at RT |
Calibration | NIBSC (93/730) value (IU/mL) = 0,7 x HTRF mIL6 value (pg/mL) |
Species | Mouse only |
Intra-assay (n=24)
Sample | Mean [IL6] (pg/mL) | CV |
---|---|---|
1 | 40 | 12% |
2 | 1400 | 7% |
3 | 2800 | 11% |
Mean CV | 10% |
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 | 72 | 503 | 6% |
2 | 448 | 3188 | 10% |
3 | 2800 | 18091 | 10% |
Mean CV | 9% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
This guide provides you an overview of HTRF applications in several therapeutic areas.
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