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HTRF Human IL-12 Detection Kit, 500 Assay Points
The HTRF human IL12 kit is designed for the quantification of IL12 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).
Product Variants
Product information
Overview
Also known as NKSF (natural killer cell stimulatory factor) and CLMF (cytotoxic lymphocyte maturation factor), IL12 is produced by macrophages and B cells. Along with IFN gamma, IL12 promotes the differentiation of naïve T cells into TH1 cells, which in turn release IFNg and IL2. Because of its effects on TH1 induction, IL12 is strongly linked to autoimmunity. IL12 is a heterodimer of p35 and p4,0 which is shared with IL23. Revvity's IL12 assay is specific for IL12p70 (full form) and does not cross react with IL23.
Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma.
Specifications
| Assay Points |
500
|
|---|---|
| Assay Target Type |
Kit
|
| Assay Technology |
HTRF
|
| Brand |
HTRF
|
| Quantity |
1
|
| Therapeutic Area |
Oncology & Inflammation
|
| Unit Size |
500 Assay Points
|
Video gallery
Citations
How it works
Assay principle
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.
Assay data analysis
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.
Assay details
Technical specifications of human IL12 kit
| Sample size | 16 µL |
|---|---|
| Final assay volume | 20 µL |
| Kit components | Lyophilized standard, frozen detection antibodies, buffers &protocol. |
| LOD &LOQ (in Diluent) | 14 pg/mL &81 pg/mL |
| Range | 81 13,000 pg/mL |
| Time to result | 2h at RT |
| Calibration | NIBSC (95/544) value (IU/mL) = 0,01 x HTRF hIL12 value (pg/mL) |
| Species | Human only |
Analytical performance
Intra and inter assay
Intra-assay (n=24)
| Sample | Mean [IL12] (pg/mL) | CV |
|---|---|---|
| 1 | 200 | 6% |
| 2 | 5000 | 4% |
| 3 | 10000 | 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 | [IL12] (pg/mL) | Mean (delta R) | CV |
|---|---|---|---|
| 1 | 252 | 444 | 8% |
| 2 | 1221 | 2158 | 11% |
| 3 | 5909 | 10789 | 12% |
| Mean CV | 10% |
Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.
Resources
Guide
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SDS, COAs, Manuals and more
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