Western blot analysis was performed on recombinant proteins, 20 ng of IL-6 (Lane 1) and 50 ng of IL-6 (Lane 2). The blots were probed with ABfinity™ Rabbit Monoclonal Anti-IL6 Antibody (Product# 701028, 0.1-1 ug/ml) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate(Product # G21234,1:5000 dilution). A 21 kDa band corresponding to IL6 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX),XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001).The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependant|
|Flow Cytometry (Flow)||1:100|
|Western Blot (WB)||0.1-1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|ELISA (ELISA)||See 1 publications below|
701028 was successfully to detect IL-6 in Flow application.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Interleukin 6 is a cytokine secreted by T cells and macrophages, has shown to exhibit activity in response to tissue damage and infection. They have also been found to be involved in various other activities like development of blood, cancer, embryonic development, and regulation of the immune system. IL6 is included in a family of other cytokines which include ciliary neurotrophic factor, cardiotrophin-1, leukemia inhibitory factor and interleukin 11. IL-6 can bring about differentiation through a receptor system which requires a specific receptor and a signaling subunit, gp130.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Four assay designs and on-chip calibration: gadgets for a sepsis protein array.
701028 was used in ELISA to develop four protein microarray assays for the early diagnosis of sepsis based on the measurement of CRP, PCT and IL-6
|Buchegger P,Preininger C||Analytical chemistry (86:3174)||2014|