Human IL-6 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-6 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-6 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-6 is added to the wells and binds to the combination of capture antibody- IL-6 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IL-6 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-6 standard dilutions and IL-6 sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Interleukin 6(IL-6) is a multifunctional protein produced by lymphoid and non-lymphoid cells and by normal and transformed cells, including T cells, monocyte/macrophages, fibroblasts, hepatocytes, vascular endothelial cells, cardiac myxomas, bladder cell carcinomas, myelomas, astrogliomas and glioblastomas. The production of IL-6 in these cells is regulated, either positively or negatively, by a variety of signals including mitogens, antigenic stimulation, lipopolysaccharides, IL-1, TNF, PDGF and viruses. For reviews on IL-6.

The human IL-6 cDNA sequence predicts a protein of 212 amino acid (aa) residues in length with two potential N-glycosylation sites. The hydrophobic N-terminal 28 aa residue signal peptide is cleaved to produce a mature protein of 184 amino acids with four cysteine residues and a predicted molecular mass of 21 kDa. The mouse IL-6 cDNA sequence shows a homology of 42% at the aa level when compared with the human sequence. On the basis of sequence similarity and gene structural motif similarity, IL-6 can be grouped in a family of cytokines that also includes OSM, G-CSF, LIF, and CNTF. All of these cytokines are predicted to have a four helix bundle structure similar to that found for growth hormone, suggesting that they all evolved from a common ancestral gene.

The effects of IL-6 on different cells are numerous and varied. The effect on B cells is stimulation of differentiation and antibody secretion. IL-6 also affects T cells, acting as a co-stimulant with sub-optimal concentrations of PHA or Con A to stimulate IL-2 production and IL-2 receptor expression.

IL-6 exhibits growth factor activity for mature thymic or peripheral T-cells and reportedly enhances the differentiation of cytotoxic T-cells in the presence of IL-2 or IFN-γ. IL-6 stimulates production of acute phase proteins by hepatocytes and has colony-stimulating activity on hematopoietic stem cells. IL-6 has growth factor activities and will stimulate the growth of myeloma/hybridoma/ plasmacytoma cells, EBV-transformed B cells, keratinocytes and mesangial cells.

GENE ID	3569
SWISS PROT	P05231
SYNONYMS	CDF; HGF; HSF; BSF2; IL-6; BSF-2; IFNB2; IFN-beta-2

Materials Supplied

Kit Components	96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human IL-6 (8﹡12)	1 plate
Human IL-6 Standard lyophilized, 1000 pg/ml upon reconstitution	2 vials
Concentrated Biotin-Conjugate anti-human IL-6 monoclonal antibody	2 vials
Streptavidin-HRP solution	2 vials
Standard /sample Diluent	1 bottle
Biotin-Conjugate antibody Diluent	1 bottle
Streptavidin-HRP Diluent	1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)	1 bottle
Substrate Solution	1 vial
Stop Solution	1 vial
Adhesive Films	4 pieces
Product data sheet	1 copy

Storage

Store at 2 - 8°C

Performance Characteristics

REPEATABILITY	The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY	The minimum detectable dose was 2pg/mL.
ASSAY RANGE	3.90 - 250 pg/mL
SPECIFICITY	This assay recognizes both natural and recombinant human IL-6. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.

Factors assayed for cross-reactivity

Recombinant human	CD40 Ligand, CT-1, CTLA-4, G-CSF, sgp130, IL-6 sR, IL-6 sR/sgp130, IL-9, IL-10, IL-12, IL-12/23 p40, LIF, LIF R, OSM
Recombinant mouse	IL-6, IL-10, IL-11, IL-12, IL-12/23 p40, IL-13
Recombinant rat	CNTF, G-CSF, IL-6, OSM

Data Analysis Assistance

We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKH1007.

https://www.myassays.com/bioss-human-il-6-elisa-kit.assay