Description
Key facts
Detection method:Colorimetric,
Sample types:Suspension cells,
Reacts with:Human,
Assay type:Sandwich (qualitative),
Assay Platform:Microplate
Product details:
Intended use
This Human Perforin ELISPOT Kit (with un-coated plates) 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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Perforin also referred to as PRF1 is a pore-forming protein that plays an important role in immune response. It appears as a 67-kDa protein and is mainly expressed in cytotoxic T cells and natural killer (NK) cells. The function of perforin involves the formation of transmembrane pores in the membranes of target cells aiding in the delivery of pro-apoptotic molecules. This mechanism is essential for the destruction of virus-infected cells and tumor cells. Techniques like perforin IHC and perforin ELISA are suitable for detecting and quantifying this protein in various samples.
Biological function summary
Perforin helps the immune system respond effectively against infected or malignant cells. It works in conjunction with granzymes forming a complex that perforates target cell membranes allowing granzymes to enter and induce apoptosis. Perforin's role in immune surveillance prevents the propagation of harmful cells within the body. Deficiency or malfunction in perforin can lead to compromised immune function or immune cell dysregulation.
Pathways
Perforin integrates into the cytotoxic T lymphocyte (CTL) and NK cell-mediated cytotoxicity pathways. These pathways include critical interactions with proteins like granzymes and Fas ligand (FasL) which orchestrate the apoptosis of abnormal cells. Through these pathways perforin ensures the body's ability to identify and eliminate threats. The granule exocytosis pathway in particular heavily depends on perforin's activity to execute effective immune responses.
Mutations or defects in perforin are linked to familial hemophagocytic lymphohistiocytosis (FHL) and some types of cancer. These conditions arise when perforin's ability to function correctly is impaired. For instance in FHL defective perforin hampers the immune cells' ability to kill target cells leading to uncontrolled immune activation. Furthermore in the context of cancer the proper functioning of perforin alongside proteins like granzyme B is critical for the surveillance and elimination of tumor cells.
