Human Perforin ELISA Assay

$560.00

The Human Perforin ELISA Assay is to be used for the in-vitro quantitative determination of Perforin in supernatant, buffered solutions. The assay will recognize natural Perforin. The Eagle Biosciences Human Perforin ELISA Assay kit is for Research Use Only.

SKU: PFN31-K01 Categories: , ,

Human Perforin ELISA Assay

The Human Perforin ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 40 pg/ml
Dynamic Range: 62.5 – 2000 pg/ml
Incubation Time: 2 h 45 min
Sample Type: Supernatant, Buffered Solutions
Sample Size: 100 µl
Note for serum & plasma quantification: High and non-linear detection level is found in human serum and plasma samples. The high signal appears not dependent to the antibody pairs but probably to the matrix and/or Perforin interaction with other molecules. Consequently serum and plasma quantification is under the scientist’s responsibility and specific investigations. Remark: no detection signal was found in fetal calf serum, rat, mouse and horse serum.


Assay Background

Perforin is a 70kDa protein containing 534 amino acid.

Perforin is involved in the killing function by CTLs and NKs. Upon degranulation, perforin inserts itself into the target cell’s plasma membrane, forming a pore. It leads to osmotic lysis of the target cells and subsequently enables granzymes to enter the target cells and activate apoptosis, the cell death program.

Perforin has structural and functional similarities to complement component 6, 7, 8 and 9 (C6, C7, C8, 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.

Its role in the immune response against tumors and virus infection has been demonstrated.


Related Products

Human MICA ELISA Assay Kit
Human CD138 /SYNDECAN-1 ELISA Assay Kit
Human sCD141 ELISA Assay

Additional Information

Assay Principle


The Eagle Biosciences Human Perforin ELISA Assay Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for Perforin has been coated onto the wells of the microtiter strips provided. Samples, including standards of known Perforin concentrations and unknowns are pipetted into these wells.
During the first incubation, the Perforin antigen is added to wells. After washing, a biotinylated monoclonal antibody specific for Perforin is incubated. Then the enzyme (streptavidin-peroxydase) is added. After incubation and washing to remove all unbound enzyme, a substrate solution which acts on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of Perforin present in the samples.

  1. Add 100µl of each, Sample and standard in duplicate to appropriate number of wells
  2. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour(s)
  3. Remove the cover and wash the plate
  4. Add 50µl of diluted biotinylated anti-Perforin to all wells
  5. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour(s)
  6. Repeat wash step 3.
  7. Add 100μl of Streptavidin-HRP solution into all wells
  8. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  9. Repeat wash step 3.
  10. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  11. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil.
  12. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


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