PeliKine compact human IL-6 kit

The PeliKine compact human IL-6 ELISA Kit is the ideal solution for sensitive detection of the pro-inflammatory cytokine Interleukin 6 (IL-6), which is produced by human monocytes when exposed to endotoxins or pyrogens.

Each kit contains sufficient reagents for 288 tests (including calibration curves)

Article number	M1916
Product group	Cytokines Immune reagents
Technique	ELISA

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Product information

General information

The PeliKine compact human IL-6 kit employs a ‘sandwich-type’ enzyme immunoassay format. In this method, a monoclonal antibody is pre-coated onto polystyrene microtiter wells. The target cytokine, found in a specified volume of sample or standard, is captured by the antibody on the microtiter plate. Any unbound substances are washed away. Next, a biotinylated antibody specific to the cytokine is added, which adheres to the cytokine-antibody complex in the well. After removing any excess biotinylated antibody through washing, horseradish peroxidase (HRP) conjugated to streptavidin is introduced. This enzyme binds to the biotin on the cytokine sandwich. Following another washing step to remove unbound HRP conjugate, a substrate solution is applied to the wells. The resulting colored product develops in proportion to the cytokine concentration in the sample or standard. The reaction is halted by adding a stop solution, and the absorbance is then measured at 450 nm using an absorbance reader.

Studies have shown that in kidney transplant recipients, the presence of IL-6 in serum and urine signals the start of rejection episodes. Moreover, increased IL-6 levels have been detected in the serum of patients suffering from septic shock, multiple myeloma, and alcoholic hepatitis, with significant variances noted between survivors and non-survivors. Traditionally, IL-6 has been measured using bioassays that rely on the proliferation of B-cell hybridomas. While these tests are sensitive, they are also slow and can be affected by other substances. The PeliKine compact IL-6 ELISA kit has been introduced to enable quicker, more reliable, and specific measurement of human IL-6 in plasma, other bodily fluids, and cell-culture supernatants.

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Test principle and procedure

The unique concept of the PeliKine compact kit delivers excellent performance at an attractive price.

The HPE (High Performance ELISA) dilution buffer ensures linear dose-response curves across various matrices. High sensitivities are achieved using poly-HRP (a polymer of HRP conjugated to streptavidin). Both the HPE buffer and poly-HRP are available separately.

The compact human cytokine ELISA kit includes all essential components for 288 tests, excluding buffers and enzyme substrates, which are commonly available in research laboratories.

For added convenience, these additional buffers are available as the PeliKine tool set (M1980). After an overnight coating, the assay can be completed in 4 hours. The optimized and easy-to-follow instructions for use (IFU) included in all kits ensure reliable assay results.

Storage.

Avoid repeated freeze-thawing of the standard, although experimental data have shown that up to 3 freeze-thaw cycles have no effect on the IL-6 levels of the standard.

Package contents

The PeliKine compact human IL-6 ELISA Kit contains all necessary reagents and buffers to perform 288 tests (3 ELISA plates).

Components:
Coating antibody
Blocking reagent
Cytokine standard
Biotin-conjugated anti-human cytokine antibody
Streptavidin-poly-HRP conjugate
HPE (High Performance ELISA) dilution buffer
Three microtiter plates.

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Quantity	Kit component	Volume	Cap colour
1 vial	coating antibody 100-fold concentrated	375 µl	red
1 vials	blocking reagent 50-fold concentrated	2 ml	transparent
1 vial	IL-6 standard see label	750µl	black
1 vial	biotinylated antibody 100-fold concentrated	375 µl	yellow
1 vial	streptavidin-poly-HRP conjugate 10,000-fold concentrated	20 µl	brown
1 bottle	HPE-dilution buffer 5-fold concentrated	55 ml	M2940
3 pcs	microtiter plate + lid	–	M1821
10 pcs	plate seals	–	M1823

Background

A recombinant huIL-6 standard has been calibrated against the WHO First International Standard (IL-6 89/548; National Institute for Biological Standards and Control, Potter Bar, Hertfordshire, U.K. 1 WHO Unit = 10 pg IL-6, see ref [20]. The kit contains one black-capped vial with 4000 pg/ml recombinant huIL-6.

Standard curve

Label 7 tubes, one tube for each dilution: 450, 150, 50, 16.7, 5.6, 1.9 and 0.6 pg/ml. Pipette 497 µl of working-strength dilution buffer into the tube labelled 450 pg/ml and 400 µl of workingstrength dilution buffer into the other tubes. Transfer 63 µl of the IL-6 standard (4000 pg/ml) into the first tube labelled 450 pg/ml, mix well and transfer 200 µl of this dilution into the second tube labelled 150 pg/ml. Repeat the serial dilutions five more times by adding 200 µl of the previous tube of diluted standard to the 400 µl of dilution buffer. The standard curve will contain 450, 150, 50, 16.7, 5.6, 1.9, 0.6 and 0 pg/ml (dilution buffer). It is recommended to prepare two separate series for each assay.

Samples

It is recommended to dilute the test samples at least 1:2 in working-strength dilution buffer. If high levels of IL-6 (outside the standard curve) are expected in the test samples, additional dilutions of sample i.e. 1:10 and 1:100 should also be prepared