Invitrogen™ Human Perforin ELISA Kit

Description

ELISA

The Perforin Human ELISA Kit quantitates Hu Perforin in human serum, plasma, or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu Perforin.

Principle of the method

The Human Perforin solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Perforin is one of the major cytolytic proteins of cytolytic granules. Perforin is a cytolytic mediator and is stored in and released by cytoplasmic granules. Moreover, perforin is involved in immune defense against tumors and virus infections as mediated by cytotoxic lymphocytes. Perforin is a 555 amino acid protein with a 21 amino acid signal peptide, and has a molecular weight of 70 to 75 kD. Perforin is a pore forming protein with a mechanism of transmembrane channel formation similar to C9, and homology between perforin and C9 have been demonstrated. Studies show that perforin is expressed only in killer cell lines and not in helper T lymphocytes or other tumor cells tested. Perforin is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis. Defects in the perforin 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 of perforin.

Specifications

• Accession Number: P14222
• Assay Range: 28.1 to 1800 pg/mL
• Assay Sensitivity: 11.8 pg/ml
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Cell Lysates, Plasma, Serum
• Target Species: Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 5551
• Gene Symbol: PRF1
• Interassay CV: 9.3%
• Intraassay CV: 4.9
• Kit Contents: Pre-coated 96 well plate, Standard (2 vials), Sample Diluent, Assay Buffer concentrate, Biotinylated Detection Antibody, Streptavidin-HRP, Wash Buffer, Substrate Solution, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Quantity: 10 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate 20 μL, Plasma 20 μL, Serum 20 μL
• Storage Requirements: 2°C to 8°C
• Target: Perforin
• Test Time: 1 hrs 20 mins
• Total Assay Time: 3 hrs 30 mins