Abcam, ab46068, Human Perforin ELISA Kit (PRF1)

Size: 1 x 96Tests / 2 x 96Tests
Human Perforin ELISA Kit (PRF1) is a sandwich ELISA designed to quantify Human Perforin (PRF1) with a sensitivity of 40 pg/mL. - Colorimetric sandwich ELISA - 450 nm readout - works on any plate reader - Wide dynamic range - quantifies 62.5 - 2000 pg/mL - Cited in over 10 publications
Key facts
Detection method:Colorimetric,
Sample types:Cell culture supernatant,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:< 40 pg/mL,
Range:62.5 - 2000 pg/mL,
Assay time:2h 45m,
Assay Platform:Microplate

Product details:
Human Perforin ELISA Kit (PRF1) ab46068 is a sandwich ELISA to measure Human Perforin (PRF1) in cell culture supernatant with a sensitivity of 40 pg/ml.
How the assay works
A monoclonal antibody specific for Perforin has been coated onto the wells of the microtiter strips provided. Samples, including standards of known Perforin concentrations, control specimens or unknowns are pipetted into these wells. During the first incubation, the standards or samples and a biotinylated monoclonal antibody specific for Perforin are simultaneously incubated. After washing, the enzyme Streptavidin-HRP, that binds the biotinylated antibody is added, incubated and washed. A TMB substrate solution is added which acts on the bound enzyme to induce a colored reaction product. The intensity of this colored product is directly proportional to the concentration of Perforin present in the samples.
Assay Specificity
Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details
Human Perforin ELISA Kit (PRF1) ab46068 protocol summary
1. Add standard or sample to each well used
2. Add prepared Biotinylated labeled detector antibody. Incubate at room temperature
3. Aspirate and wash each well
4. Add prepared Streptavidin-HRP mix to each well. Incubate at room temperature
5. Aspirate and wash each well.
6. Add the TMB Solution to each well until color develops and then add the Stop Solution
7. Immediately begin recording the color development

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.