IL-6 ELISA | BIOZOL

Article Name:	IL-6 ELISA
Biozol Catalog Number:	BI-IL6
Supplier Catalog Number:	BI-IL6
Alternative Catalog Number:	BI-IL6
Manufacturer:	Biomedica
Category:	Kits/Assays

BIOMEDICA´s human Interleukin-6 (IL-6) ELISA Assay is highly sensitive and shows detectable values in serum and in plasma samples.

Background

Interleukin-6 (IL-6) is a multifunctional cytokine that plays an important role in the body´s response to tissue injury or infection. Il-6 is produced by various cells, including T-cells, B-cells, macrophages, as well as vascular endothelial cells, mast cells, and dentritic cells. Il-6 secretion is stimulated during inflammatory response and travels through the bloodstream to the liver, where it induces the production of acute phase reactants such as C-reactive protein and others.

Product Overview

The Biomedica human Interleukin-6 ELISA (IL-6) ELISA kit is a sandwich enzyme immunoassay that has been optimized and fully validated for the quantitative determination of human IL-6 in serum and plasma (EDTA, heparin, citrate). Validation experiments have been performed according to international quality guidelines (ICH/ FDA/ EMEA). Cell culture supernatant and urine samples are compatible with this ELISA. The IL-6 ELISA assay recognizes both natural and recombinant human IL-6. The assay employs highly purified epitope mapped antibodies as well as human serum-based standards and controls.

Features & Benefits

High Specificity – using characterized epitope-mapped antibodies
High Sensitivity – measurable values in both serum and plasma
Superior Quality – validated according to international quality guidelines
Standardized – use of WHO intern.standards for kit calibration & harmonization
User-Friendly – ready to use color coded prediluted standards & controls

Human IL-6 ELISA Assay Principle

The figure below explains the principle of the human IL-6 sandwich ELISA:

Capture antibody: recombinant anti-human IL-6 antibody
Detection antibody: polyclonal anti-human IL-6 antibody, Biotin-labeled
Target antigen: human IL-6

This kit is a sandwich enzyme immunoassay for the quantitative determination of human IL-6 (Interleukin-6) in human serum, plasma, cell culture supernatants, and urine samples. In a first step, STD/sample/CTRL are pipetted into the wells, which are pre-coated with the recombinant anti-human IL-6 antibody. Any soluble IL-6 present in the STD/sample/CTRL binds to the pre-coated anti-IL-6 antibody in the well. After incubation, a washing step is applied where all non-specific unbound material is removed. In a next step, the biotinylated anti-IL-6 antibody (AB) is pipetted into the wells and reacts with the IL-6 present in the sample, forming a sandwich. Next, all unbound antibody is removed during another washing step. In the next step, the conjugate (streptavidin-HRPO) is added and reacts with the biotinylated anti-IL-6 antibody. After another washing step, the substrate (Tetramethylbenzidine; TMB) is pipetted into the wells. The enzyme catalysed colour change of the substrate is directly proportional to the amount of IL-6 present in the sample. This colour change is detectable with a standard microtiter plate ELISA reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated, using the values obtained from the standards. The concentration of soluble IL-6 in the sample is determined directly from the dose response curve.

Typical Data

This standard curve and the displayed OD values are for demonstration only. A standard curve should be generated for each assay run.

Standard	IL-6 [pg/ml]	OD			CV [%]
Standard	IL-6 [pg/ml]	#1	#2	Average	CV [%]
STD1	0	0.056	0.059	0.058	4
STD2	3.125	0.140	0.129	0.135	6
STD3	6.25	0.216	0.207	0.212	3
STD4	12.5	0.354	0.366	0.360	2
STD5	25	0.661	0.660	0.661	0
STD6	50	1.090	1.138	1.114	3
STD7	100	1.970	1.835	1.903	5
STD8	200	2.951	2.894	2.923	1

The quality control protocol supplied with the kit shows the results of the final release QC for each kit at the production date. ODs obtained by customers may differ due to various influences including a normal decrease of signal intensity throughout shelf life. However, this does not affect the validity of the results, provided an OD of 1.50 or higher is obtained for the standard with the highest concentration, and the measured control values fall into their respective target range (see labels).

IL-6 ELISA Kit Components

All reagents supplied in the human IL-6 ELISA kit are stable at 2-8°C until the expiry date stated on the label of each reagent. Reconstituted reagents must be stored at -25°C until the expiry date.

CONTENT	DESCRIPTION	QUANTITY
PLATE	Microtiter strips coated with recombinant IL-6 antibody specific for human IL-6 in strip holder packed in an aluminum bag with desiccant	12 x 8 tests
WASHBUF	20x wash buffer concentrate, transparent cap	1 x 50 ml
ASYBUF	Assay buffer, red cap, ready to use	1 x 10 ml
STD	Recombinant IL-6 standards (0 / 3.125 / 6.25/ 12.5 / 25 / 50 / 100 / 200 pg/ml), human serum based, white caps, lyophilized	8 vials
CTRL	Control A and B, human serum based, yellow cap, lyophilized, exact concentration is stated on labels	2 vials
AB	Polyclonal IL-6 antibody specific for human IL-6, biotin-labeled, green cap, ready to use	1 x 13 ml
CONJ	Conjugate (streptavidin-HRPO), brown cap, ready to use	1 x 13 ml
SUB	Substrate (TMB solution), blue cap, ready to use	1 x 13 ml
STOP	STOP solution, white cap, ready to use	1 x 7 ml

Product Details

Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
Sensitivity LOD: 0.28 pg/ml (measurable concentrations in serum AND plasma samples!)
Sample types: human serum, plasma (EDTA, citrate, heparin), cell culture supernatants, urine)
Sample volume: 100 µl/well
Dynamic range: 0 – 200 pg/ml (0 / 3.12 / 6.25 / 12.5 / 25/ 50 / 100 / 200)
Time to result: 4.5 hours
Specificity: Endogenous and recombinant human IL-6
Protokol booklet
Validation data file
Alternative name: Interleukin-6

This typical data standard curve and the displayed OD values are for demonstration only. A standard curve should be generated for each assay run.