High-sensitivity IL-6 Products
IL-6 is one kind of interleukins which exerts pro and anti-inflammatory effects in the body. It’s a typical pro-inflammatory cytokine and encoded by IL-6 gene. T cell and macrophage will been differentiated into activated cells and proliferate when outer substances encounted, then much IL-6 was produced to stimulate immune responses. The level of IL-6 increases significantly in infection, wound, burn and other tissues lesions. It is up-regulated by IL-1ra and IL-10 and down-regulated by TNF-alpha and IL-1. In clinical research, it has been found that IL-6 is associated with many systematic diseases, such as periodontitis, acute myocardial infarction, gestational hypertension, prostatic carcinoma and tumors.
With the increasing studies of IL-6, a series of IL-6 detection tools were been developed, which include recombinant proteins, antibodies and Elisa kits. As the basal level of IL-6 is relatively low in the body, we developed high sensitivity IL-6 Elisa kits, and commercialized the monoclonal antibody with high purity and specificity in the kit.
The human IL-6 protein has 212 amino acids, the molecular weight is about 24 kDa. As the first 29 amino acids at the N-terminal are a signal peptide, then we chose Pro29~Met212 to construct plasmid and express this construct in E.coli, then we got recombinant protein and purified it by affinity column chromatography (RPA079Hu01). As there is a GST tag on the protein, the MW is about 47 kD. Finally, we verified its molecular weight and purity by SDS-PAGE, as shown in Fig.1.
Immunizing rabbit with RPA079Hu01 and purifying the polyclonal antibodies with antigen coupling affinity column chromatography and we get high purity rabbit anti-human IL-6 polyclonal antibodies(PAA079Hu01).Coupling PAA143Hu01 with biotin to get biotin coupled polyclonal antibodies(PAA079Hu71), which bring much access to detect IL-6 in vivo and in vitro. Western Blot was gone with PAA079Hu01 to detect plasma samples, please see Fig 2. IHC experiments were carried with human liver, it shows the IL-6 is mainly secrected in liver.
On the development of IL-6 recombinant protein, antibodies, Cloud-Clone Corp. developed IL-6 detection kits, including Elisa kit(SEA079Hu)and CLIA kit(SCA079Hu).
MAA079Hu22) by immunizing mouse with RPA079Hu01 and purifying with affinity column chromatography. It needs minim and with high sensitivity on SDS-PAGE, WB and IHC experiments.
On the basis of high purity and specificity monoclonal antibodies, the high sensitivity kit was developed by repeatedly adjustment. The sensitivity means the concentration of mean OD of 20 blank (the standard diluent) plus two times standard deviation. The minimum detection limit is 0.156 pg/ml and the sensitivity is 0.047pg/ml of high sensitivity kit; which is 10 times compared to the minimum detection limit and the sensitivity in normal kit, respectively.
Besides human IL-6 products, Cloud-Clone Corp. successfully developed about 10 species includes rat, mouse, rabbit, dog, etc. and over 70 IL-6 detection products, including proteins, antibodies, Elisa kits, CLIA kits. We also have high sensitivity kit on rattus norvegicus, these products from different species provide perfect technical support to study the expression and function of IL-6.