MAP-Tau standard
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
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Highly specific
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Faster and more convenient than ELISA
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.
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.
intra and inter assay
Intra-assay (n=24)
| Sample | Mean [Progranulin] (pg/mL) | CV | ||
|---|---|---|---|---|
| 1 | 781 | 2.1% | ||
| 2 | 3,125 | 2.1% | ||
| 3 | 6.250 | 1.4% | ||
| 4 | 25,000 | 1.2% | ||
| 5 | 50,000 | 1.2% | ||
| 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)
| Sample | Mean [Progranulin] (pg/mL) | CV | ||
|---|---|---|---|---|
| 1 | 777 | 11% | ||
| 2 | 1,787 | 12% | ||
| 3 | 4,109 | 13% | ||
| 4 | 21,739 | 10% | ||
| 5 | 50,000 | 8% | ||
| 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 | - |
| 2 | 21,301 | 21,100 | 99% |
| 4 | 10,650 | 10,524 | 99% |
| 8 | 5,325 | 5,023 | 94% |
| 16 | 2,663 | 2,711 | 102% |
| 32 | 1,331 | 1,398 | 105% |
| 64 | 666 | 707 | 106% |
| Mean | 101% | ||
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 | 492 | 1,911 | 1,788 | 97% | |
| 2,199 | 3,618 | 3,339 | 92% | ||
| 11,139 | 12,558 | 12,187 | 97% | ||
2 4,440 pg/mL | 492 | 4,932 | 4,850 | 98% | |
| 2,199 | 6,639 | 6,285 | 95% | ||
| 11,139 | 15,579 | 16,201 | 104% | ||
3 19,493 pg/mL | 492 | 19,985 | 19,703 | 99% | |
| 2,199 | 21,692 | 21,490 | 99% | ||
| 11,139 | 30,632 | 32,267 | 105% |
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)
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
Product Insert Progran (h) Kit 2 / 62PROPEG-62PROPEH
62PROPEG-62PROPEH - Product Insert
Safety Data Sheet (DEU) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (ELL) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (ITA) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (SPA) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) Progran (h) Kit 2 / 62PROPEG
62PROPEG - Safety Data Sheet
Safety Data Sheet (DEU) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (ELL) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (ITA) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (SPA) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) Progran (h) Kit 2 / 62PROPEH
62PROPEH - Safety Data Sheet
Plate Reader Requirement
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