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HTRF Human Progranulin Detection Kit, 500 Assay Points
HTRF Human Progranulin Detection Kit, 500 Assay Points
Generic HTRF biomarker image
The Progranulin HTRF kit is designed for the accurate quantitative measurement of Progranulin produced by cells.
| Feature | Specification |
|---|---|
| Application | タンパク質定量 |
| Sample Volume | 16 µL |
The Progranulin HTRF kit is designed for the accurate quantitative measurement of Progranulin produced by cells.
Product variants
Unit Size: 500 assay points
Part #:
62PROPEG
Unit Size: 10,000 assay points
Part #:
62PROPEH
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).
HTRF Human Progranulin Detection Kit, 500 Assay Points
Generic HTRF biomarker image
HTRF Human Progranulin Detection Kit, 500 Assay Points
Product information
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.
HTRF assays offer many advantages over other technologies:
- Homogeneous add-and-read format
- No wash steps
- Low background
- Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
- Stable signal, providing flexibility in time of readout or size of assays
How it works
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.
Assay details
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% |
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% |
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.
Assay validation
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)
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
|
| Unit Size |
500 assay points
|
Video gallery
HTRF Human Progranulin Detection Kit, 500 Assay Points
HTRF Human Progranulin Detection Kit, 500 Assay Points
Resources
Are you looking for resources, click on the resource type to explore further.
Guide
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