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Human Progranulin kit HTRF®

The Progranulin HTRF kit is designed for the accurate quantitative measurement of Progranulin produced by 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 Progranulin HTRF kit is designed for the accurate quantitative measurement of Progranulin produced by cells.
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Overview

Progranulin, also called proepithelin or prostate cancer (PC) cell-derived growth factor, is the precursor protein of granulin. Progranulin is involved in Frontotemporal Lobar Degeneration (FTLD) and Alzheimer's Disease, where it may protect against beta amyloid toxicity. This cell-based HTRF assay ensures quick results, and is adapted to the functional screening of compounds using progranulin secreting cells.

Benefits

  • CELL AND CELL SUPERNATANT FORMATS
  • LOW SAMPLE CONSUMPTION

Assay principle

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

Assay Protocol

The simple Progranulin assay protocol, using a 384-well small volume white plate (20 µL final), is described on the right. Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection of human Progranulin. The antibodies labelled with the HTRF donor and acceptor may 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.

Human progranulin kit assay protocol

intra and inter assay

Intra-assay (n=24)

SampleMean [Progranulin] (pg/mL)CV
17812.1%
23,1252.1%
36.2501.4%
425,0001.2%
5​​​​​​​50,0001.2%

Mean CV
1.6%

Each of the 5 samples was measured 24 times, and the % CV was calculated for each sample.

The sample used was a supernatant collected on unstimulated HMC3 cells.

Inter-assay (n=4)

SampleMean [Progranulin] (pg/mL)CV
177711%
21,78712%
34,10913%
421,73910%
5​​​​​​​50,0008%

Mean CV
11%

Each of the samples was measured in 4 different experiments, and the % CV was calculated for each sample.

Dilutional linearity

Dilution factor[Progranulin] expected (pg/mL)[Progranulin] detected (pg/mL)Recovery
1-42,602-
221,30121,10099%
410,65010,52499%
85,3255,02394%
162,6632,711102%
321,3311,398105%
64666707106%
Mean101%

The excellent recovery % obtained from these experiments show the good linearity of the assay.

Spike and recovery

Sample

[Progranulin] added (pg/mL)[Progranulin] expected (pg/mL)[Progranulin] detected (pg/mL)Recovery

1

1,419 pg/mL

4921,9111,78897%
2,1993,6183,33992%
11,13912,55812,18797%

2

4,440 pg/mL

4924,9324,85098%
2,1996,6396,28595%
11,13915,57916,201104%

3

19,493 pg/mL

49219,98519,70399%
2,19921,69221,49099%
11,13930,63232,267105%

​The same amount of recombinant protein was added to 3 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery found validates the sample matrix used for this assay.

Progranulin secretion from iPSC-microglia

iPSC-Microglia were derived as in McQuade et al. 2018. iPS-Microglia were stimulated with 10 ng / mL cytokines for 24 hours in the absence of TGF-b1. Cell supernatants were collected, and secreted levels of Progranulin were measured following the procedure given in the HTRF kit package insert. In agreement with the literature, IFN-γ + IL-1b (pro-inflammatory) treated microglial cells displayed a decreasing progranulin release trend, whereas anti-inflammatory IL-4 and IL-13 Th2 cytokines displayed a significant increase in secreted progranulin.


Data kindly provided by Amanda McQuade and Dr. Mathew Blurton-Jones (University of California, Irvine)

Validation of the HTRF progranulin kit on the secretion from iPSC-microglia

HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Molecular basis of neuroinflammation and neurodegeneration diseases

The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides

Neurodegeneration and its main related diseases

Discover this infographic design on neurodegenerative diseases - Infographics

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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