IL10 + IL4 Human Elispot Kit (Reagents for 20 x 96 tests, without plates) (ab48749)
- Product nameIL10 + IL4 Human Elispot Kit (Reagents for 20 x 96 tests, without plates)See all IL10 + IL4 kits ...
- Tests20 x 96 well plate
- Sample typeCell culture supernatant, Cell culture extracts
- Assay typeSandwich (qualitative)
- Assay durationMultiple steps standard assay
- Species reactivityReacts with: Human
- Product overview
The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, oncology, infectious diseases, autoimmune diseases and transplantation.
The Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.
The dual colour Elispot allows you to monitor the production of two cytokines simultaneously in the same well.
After cell stimulation, locally produced cytokines are captured by IL10 and IL4 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IL10 FITC conjugated antibody and a biotinylated anti-IL4 antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate IL10 production while IL4 is revealed by blue/purple spots.
- Tested applicationsMeasuring secreted proteins from cells more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 20 x 96 tests 10 x concentrate buffer for the preparation of AEC buffer 4 x 5ml 50 x concentrate AEC substrate buffer 4 x 1ml Anti-FITC antibody HRP conjugate 4 x 100µl Bovine Serum albumin 4 x 1g Human IL-10 Capture antibody 4 x 0.5ml IL10 FITC conjugated detection antibody 4 x 0.55ml IL-4 Biotinylated detection antibody 4 x 0.55ml IL-4 Capture antibody 4 x 0.5ml Ready-to-use BCIP/NBT substrate buffer 4 x 50ml Streptavidin - Alkaline Phosphatase conjugated 4 x 50µl
- Research Areas
- RelevanceInterleukins (ILs) are a large group of cytokines that are produced mainly by leukocytes, although some are produced by certain phagocytes and auxiliary cells. ILs have a variety of functions, but most function to direct other immune cells to divide and differentiate. Each IL acts on a specific, limited group of cells through a receptor specific for that IL. Human IL10 is a non glycosylated polypeptide consisting of 160 amino acids. There is 73% homology between the human and mouse IL10 proteins, however, the human IL10 acts on both human and mouse target cells, while the mouse IL10 has species specific activity. The cellular sources of IL10 are CD4+ T cells and T cell clones, thymocytes, B cells and B cell lymphomas, macrophages, mast cell lines and keratinocytes. IL10 will stimulate the growth of stem cells, mast cells and thymocytes. IL10 enhances cytotoxic T cell development, and costimulates B cell differentiation and immunoglobulin secretion. IL10 inhibits cytokine production by macrophages and suppresses macrophage class II MHC expression. The human IL10 gene is on human chromosome 1. IL4 is a pleiotropic cytokine produced by activated T cells, mast cells and basophils. It is a ligand for Interleukin 4 receptor. The Interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. IL4 elicits many different biological responses, but has two dominant functions. The first is regulating differentiation of naïve CD4+ T cell to the Th2 type. Th2 cells produce IL4, IL5, IL10 and IL13, which tend to favor a humoral immune response while suppressing a cell mediated immune response controlled by Th1 cells. STAT6, a signal transducer and activator of transcription, has been shown to play a central role in mediating the immune regulatory signal of this cytokine. The second is regulating IgE and IgG1 production by B cells. Two alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported.
- Cellular localizationSecreted
- B cell stimulatory factor 1
- Interleukin 10
- Interleukin 4
Our Abpromise guarantee covers the use of ab48749 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
References for IL10 + IL4 Human Elispot Kit (Reagents for 20 x 96 tests, without plates) (ab48749)
ab48749 has not yet been referenced specifically in any publications.