Human Perforin-1 kit HTRF®

The Human Perforin-1 kit is designed for the rapid detection of human Perforin-1 in cell supernatant and whole cells.
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  • Ready-to-use Ready-to-use
  • Highly specific Highly specific
  • Faster and more convenient than ELISA Faster and more convenient than ELISA
The Human Perforin-1 kit is designed for the rapid detection of human Perforin-1 in cell supernatant and whole cells.
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Overview

Perforin (also called Cytolysin or Lymphocyte pore-forming protein) is found in cytotoxic lymphocytes, NK cells (natural killer), and cytotoxic T cells. Perforin creates a pore in the cell membrane to allow Granzyme B to enter the target cell’s cytoplasm and triggers apoptosis through caspase activation. This kit is designed for the rapid detection of human Perforin in cell supernatant and whole cells. The assay enables high throughput without any washing steps, thereby saving time compared to ELISA.

Benefits

  • LOW SAMPLE CONSUMPTION
  • CTLS, NK AND CYTOTOX T CELLS STUDY

Assay principle

Human Perforin is measured with a sandwich immunoassay involving two specific anti-human Perforin antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of Perforin present in the sample.
Perforin kit assay principle

Assay Protocol

The assay protocol, using a 384-well small volume white plate or a Cisbio low volume 96 well plate (20 µL final), is described on the right. 16 µL of cell supernatant, sample, or standard are dispensed directly into the detection plate for detection by HTRF reagents. The antibodies labelled with HTRF donor and acceptor can be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.
Perforin kit assay protocol

Key features

Sample size 16 µL
LOD &LOQ (in diluent) 0.52 ng/mL &1.05 ng/mL
Assay range 1.05 – 100 ng/mL
Time to result ON at RT
Human Perforin-1 standard curve

Detection of Perforin on cell supernatant (Two-plate-protocol)

Different cell densities of TALL-104 cells (Effector cells) were plated in suspension under 50 µL in 96-well plate. TALL-104 cells were cultured for 24 hours at 37 °C - 5% CO2. After culture, 16 µL of cell supernatants cells were transferred into a 384-well sv white microplate and 4 µL of the HTRF Perforin and Granzyme B detection reagentswere added. The HTRF signal was recorded after an overnight incubation at room temperature.
Perforin and Granzyme B detection in a two-plate -protocol

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