ab105257 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. Fluorospot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, oncology, infectious diseases, autoimmune diseases and transplantation.
This Kit is based on sandwich immune-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.
After cell stimulation, locally produced cytokines are captured by Interferon gamma and IL17A specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti- Interferon gamma ?FITC conjugated antibody and a biotinylated anti- IL17A antibody. Those are in turn recognised by anti-FITC green fluorescent dye and streptavidin-phycoerythrin conjugates. PVDF-bottomed-well plates are then read under a UV light beam. Green fluorescent spots indicate Interferon gamma ?production while IL17A is revealed by red spots. Yellow spots will indicate dual cytokine producing cells.
See Figure 1 for visual scheme.
ab105257 provides enough reagents for 10 plates. Plates are provided, non sterile.
|Components||10 x 96 tests|
|Anti-FITC antibody green fluorescence conjugate||2 vials|
|Biotinylated detection antibody for Interferon gamma (resuspend in 0.55mL)||2 vials|
|Bovine Serum albumin||2 x 1g|
|Capture antibody for IL17A||2 x 0.5ml|
|Capture antibody for Interferon gamma||2 x 0.5ml|
|FITC conjugated detection antibody for IL17A (resuspend in 0.55mL)||2 vials|
|Fluorescence buffer||2 x 2.5ml|
|Streptavidin-phycoerythrin conjugate||2 vials|
Our Abpromise guarantee covers the use of ab105257 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|FluroSpot||Use at an assay dependent dilution.|
ab105257 has not yet been referenced specifically in any publications.