Perforin ELISpot Set | PRF1 elispot set

Human Perforin ELISpot Set

Cat. Num. AAA335726

• Gene Names: PRF1; P1; PFP; FLH2; PFN1; HPLH2
• Reactivity: Human
• Synonyms: Perforin; Human Perforin ELISpot Set; Perforin (with non-sterile plate); PRF1 elispot set

• Reactivity: Human
• Specificity: The assay recognizes natural human Perforin. To define the specificity several proteins were tested for cross reactivity. There was no cross reactivity observed for any protein tested (Granzyme B, Fas, Fas L, IL-1beta, IL-2, IFNgamma, TNFalpha and TRAIL). This testing was performed using the equivalent human Perforin antibody pair in an ELISA assay.
• Sequence Length: 555

• Intended Uses: ELISpot is a highly specific immunoassay for the analysis of cytokine and other soluble molecule production and secretion from T-cells at a single cell level in conditions closely comparable to the in-vivo environment with minimal cell manipulation. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The ELISpot assay constitutes an ideal tool in the investigation of Th1 / Th2 responses, vaccine development, viral infection monitoring and treatment, cancerology, infectious disease, autoimmune diseases and tranplantation. Utilising sandwich immuno-enzyme technology, ELISpot assays can detect both secreted cytokines and single cells that simultaneously produce multiple cytokines. 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.

Related Product Information for PRF1 elispot set

Principle of the assay: A capture antibody highly specific for the analyte of interest is coated to the wells of a PVDF bottomed 96 well microtitre plate either during kit manufacture or in the laboratory. The plate is then blocked to minimise any non-antibody dependent unspecific binding and washed. Cell suspension and stimulant are added and the plate incubated allowing the specific antibodies to bind any analytes produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured analyte. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in coloured spots which can be quantified using appropriate analysis software or manually using a microscope.

Product Categories/Family for PRF1 elispot set

Human ELISpot Set

References

Van Gulck, E. R. et al, J. Virol,2008; 82(7): 3561-3573.

NCBI and Uniprot Product Information

• NCBI GI #: 189847
• NCBI GeneID: 5551
• Molecular Weight: 61,377 Da
• NCBI Official Full Name: perforin
• NCBI Official Synonym Full Names: perforin 1 (pore forming protein)
• NCBI Official Symbol: PRF1
• NCBI Official Synonym Symbols: P1; PFP; FLH2; PFN1; HPLH2
• NCBI Protein Information: perforin-1
• UniProt Protein Name: Perforin-1
• Protein Family: Perforin
• UniProt Gene Name: PRF1
• UniProt Synonym Gene Names: PFP; P1; PFP
• UniProt Entry Name: PERF_HUMAN

NCBI Description

The protein encoded by this gene 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. This protein 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. Alternative splicing results in multiple transcript variants encoding the same protein. [provided by RefSeq, Jul 2008]

Uniprot Description

PRF1: Plays a key role in secretory granule-dependent cell death, and in defense against virus-infected or neoplastic cells. 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. Can insert into the membrane of target cells in its calcium-bound form, oligomerize and form large pores. Promotes cytolysis and apoptosis of target cells by facilitating the uptake of cytotoxic granzymes. Monomer, as sobluble protein. Homooligomer. Oligomerization is required for pore formation. Repressed by contact with target cells. Belongs to the complement C6/C7/C8/C9 family.

Protein type: Membrane protein, multi-pass

Chromosomal Location of Human Ortholog: 10q22

Cellular Component: membrane; cytoplasmic membrane-bound vesicle; integral to membrane; extracellular region; plasma membrane

Molecular Function: protein binding; calcium ion binding; wide pore channel activity

Biological Process: formation of immunological synapse; apoptosis; cytolysis; defense response to tumor cell; cellular defense response; immune response to tumor cell; defense response to virus; transmembrane transport; protein homooligomerization

Disease: Lymphoma, Non-hodgkin, Familial; Aplastic Anemia; Hemophagocytic Lymphohistiocytosis, Familial, 2