Product nameHuman Perforin ELISPOT Kit
See all Perforin kits
Sample typeCell culture supernatant, Cell culture extracts
Assay typeSandwich (qualitative)
Assay durationMultiple steps standard assay
Species reactivityReacts with: Human
This ELISPOT assay is designed to enumerate Perforin producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing perforin production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of perforin 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, cancerology, infectious diseases, autoimmune diseases and transplantation.
Abcam 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.
Principle of the method
After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells
Recognizes natural human Perforin
Tested applicationsSuitable for: ELISpotmore details
Storage instructionsStore at +4°C. Please refer to protocols.
Components 5 x 96 tests 10 x 96 tests 96 PVDF-bottomed-well plates. 5 units 10 units Bovine Serum Albumin 1 x 1g 2 x 1g Human Perforin Capture Antibody 1 x 500µl 2 x 500µl Perforin Biotinylated Detection Antibody 1 vial 1 vial Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl
RelevancePerforin has structural and functional similarities to complement component 9 (C9). Like C9, this protein creates transmembrane tubules and is capable of lysing non-specifically a variety of target cells. Perforin is one of the main cytolytic proteins of cytolytic granules, and it is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis. Defects in this gene cause familial hemophagocytic lymphohistiocytosis type 2 (HPLH2), a rare and lethal autosomal recessive disorder of early childhood.
Cellular localizationCell Membrane and Cytoplasmic
- Perforin 1
Our Abpromise guarantee covers the use of ab46620 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISpot||Use at an assay dependent dilution.|
ab46620 has not yet been referenced specifically in any publications.