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AB46620

Human Perforin ELISPOT Kit (with un-coated plates)

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Human Perforin ELISPOT Kit (with un-coated plates) is a ELISPOT kit for the sensitive detection of Human Perforin (with un-coated plates) production in a single cell suspension.

View Alternative Names

PFP, PRF1, Perforin-1, P1, Cytolysin, Lymphocyte pore-forming protein

Key facts

Detection method

Colorimetric

Sample types

Suspension cells

Reacts with

Human

Assay type

Sandwich (qualitative)

Assay Platform

Microplate

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ELISPOT": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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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
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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.
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Product protocols

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Target data

Pore-forming protein that plays a key role in granzyme-mediated programmed cell death, and in defense against virus-infected or neoplastic cells (PubMed : 20889983, PubMed : 21037563, PubMed : 24558045, PubMed : 9058810, PubMed : 9164947). Plays an important role in killing other cells that are recognized as non-self by the immune system, e.g. in transplant rejection or some forms of autoimmune disease (PubMed : 9058810). Can insert into the membrane of target cells in its calcium-bound form, oligomerize and form large pores (PubMed : 20889983, PubMed : 21037563). Promotes cytolysis and apoptosis of target cells by mediating the passage and uptake of cytotoxic granzymes (PubMed : 20038786, PubMed : 20225066, PubMed : 24558045, PubMed : 32299851). Facilitates the delivery of cationic cargo protein, while anionic or neural proteins are not delivered efficiently (PubMed : 24558045). Perforin pores allow the release of mature caspase-7 (CASP7) into the extracellular milieu (By similarity).
See full target information PRF1
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websiteProtocolBooklet
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