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  • HTRF Human Perforin-1 Detection Kit, 500 Assay Points

HTRF Human Perforin-1 Detection Kit, 500 Assay Points

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HTRF Human Perforin-1 Detection Kit, 500 Assay Points
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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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Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The Human Perforin-1 kit is designed for the rapid detection of human Perforin-1 in cell supernatant and whole cells.

View product information
Product variants
Unit Size: 500 assay points
Part #:
63ADK100PEG
Unit Size: 10,000 assay points
Part #:
63ADK100PEH
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
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Generic HTRF biomarker image
Generic HTRF biomarker image
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HTRF Human Perforin-1 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
Video Play Button
Generic HTRF biomarker image
Generic HTRF biomarker image
HTRF Human Perforin-1 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
Generic HTRF biomarker image
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Product information

  • Overview
  • How it works
  • Analytical performance
  • Assay validation
  • Specifications
  • Video gallery

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 cells 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.

How it works

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.

1biomarkers-how-it-works-assay-principle-perforin-63adk100peg-63adk100peh-63adk100pej.svg

 

Assay protocol

The assay protocol, using a 384-well small volume white plate or a Revvity 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.

2biomarkers-how-it-works-assay-protocol-perforin-63adk100peg-63adk100peh-63adk100pej.svg

 

Analytical performance

Sample size 16 µL
LOD &LOQ (in diluent) 0.1 ng/mL &0.25 ng/mL
Assay range 0.25 – 100 ng/mL
Time to result ON at RT

 

 

3assay-validation-human-perforin-std-curve.svg

 

Assay validation

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.

4assay-validation-human-perforin-1-1.svg

 

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
16 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Biomarkers
Target Species
Human
Technology
TR-FRET
Therapeutic Area
Oncology & Inflammation
Unit Size
500 assay points

Video gallery

HTRF Human Perforin-1 Detection Kit, 500 Assay Points
HTRF Human Perforin-1 Detection Kit, 500 Assay Points

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