Description
Key facts
Detection method:Colorimetric,
Sample types:Suspension cells,
Reacts with:Human,
Assay type:Sandwich (qualitative),
Assay Platform:Microplate
Product details:
This ELISPOT assay is designed to enumerate Granzyme B producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing Granzyme B production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of Granzyme B 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.
Principal:
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 Granzyme B
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.
Granzyme B also known as GZMB GB11 or granzyme B protein is a serine protease with a molecular mass of approximately 32 kDa. It is expressed mainly in cytotoxic T lymphocytes and natural killer (NK) cells. This enzyme plays a mechanical role in inducing apoptosis in target cells serving as an effector protein in the immune system's defense against virally infected cells or transformed cancer cells. The activity of granzyme B relies on its ability to cleave after aspartate residues in substrate proteins leading to the activation of apoptotic pathways.
Biological function summary
Granzyme B participates prominently in the immune response by activating caspases particularly caspase-3 which promotes the breakdown of cellular components necessary for apoptosis. Granzyme B does not function in isolation but acts in concert with other immune system factors such as perforin to effectively induce cell death. Perforin creates pores in the target cell membrane allowing granzyme B to enter and instigate the apoptosis sequence. The enzyme also contributes to the processing of cytokines which enhances the immune response further.
Pathways
Studies have determined that granzyme B is critical in the apoptosis pathway particularly in the granule exocytosis pathway. It closely interacts with proteins such as perforin and other granzymes to mediate apoptosis in target cells. Granzyme B also plays a role in the inflammatory response and can influence pathways associated with cytotoxic T cell signaling. Its pathway interactions ensure effective elimination of damaged or infected cells maintaining tissue homeostasis.
Granzyme B has associations with autoimmune diseases and cancer. Abnormally high levels of granzyme B can contribute to tissue damage and inflammation in autoimmune conditions like rheumatoid arthritis. In the context of cancer granzyme B aids in tumor surveillance and destruction when functioning correctly but impaired granzyme B activity can lead to evasion of immune detection by cancerous cells. Perforin also plays a role in these conditions closely working with granzyme B to either protect against or drive disease progression.
