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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 and disposal requirements under European REACH regulations (EC 1907/2006).

Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The HTRF human IL12 kit is designed for the quantification of IL12 release in cell supernatant.

Product Variants
Unit Size: 500 Assay Points
Part #:
62HIL12PEG
List Price
USD 1,103.00
Unit Size: 10,000 Assay Points
Part #:
62HIL12PEH
List Price
USD 10,290.00

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

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
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.

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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.

 

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