The HTRF human IL4 kit is designed for the quantification of human IL4 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).
IL4 is expressed by T cells, NK cells, and mast cells, and is an important regulator of humoral and adaptative immunity. It stimulates activated B cells, and induces ir differentiation into plasma cells and ir class switch to immunoglobulin E, as well as promoting class II MHC expression. In immuno-oncology, IL4 induces differentiation of naïve T cells into TH2 cells, which in turn produce IL4 and thus create a positive feedback loop. IL4 is also involved in development of airway inflammation and allergic asthma.
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
|
Technology |
TR-FRET
|
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) | 8 pg/mL &29 pg/mL |
Range | 29 4,000 pg/mL |
Time to result | 2h at RT |
Calibration | NIBSC (88/656) value (IU/mL) = 0,012 x HTRF hIL4 value (pg/mL) |
Species | Human only |
Intra-assay (n=24)
Sample | Mean [IL4] (pg/mL) | CV |
1 | 50 | 7% |
2 | 1895 | 2% |
3 | 3598 | 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 | [IL4] (pg/mL) | Mean (delta R) | CV |
1 | 98 | 527 | 12% |
2 | 432 | 2061 | 7% |
3 | 1905 | 7265 | 6% |
Mean CV | 8% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample
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