Amphiregulin (AR) is an EGF family growth factor that is released as a soluble protein following proteolytic cleavage of its transmembrane precursor. Amphiregulin is expressed by numerous carcinoma cell lines and epithelial cells of the colon, stomach, breast, ovary and kidney. It acts through ErbB family of receptor to stimulate the proliferation of keratinocytes, mammary epithelial cells, fibroblasts, astrocytes and glial cells. Amphiregulin can also inhibit the growth of certain tumor cells.
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Human Amphiregulin Quantikine ELISA Kit
R&D Systems | Catalog # DAR00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Saliva (50 µL), Urine (50 µL)
Sensitivity
3.56 pg/mL
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
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Product Summary for Human Amphiregulin Quantikine ELISA Kit
The Quantikine Human Amphiregulin Immunoassay is a 4.5 hour solid
phase ELISA designed to measure human Amphiregulin in cell culture
supernates, cell lysates serum, plasma, saliva, and urine. It contains E. coli-derived
recombinant human Amphiregulin and has been shown to accurately quantitate the recombinant factor.
Results obtained with natural human Amphiregulin showed
linear curves that were parallel to the standard curves obtained using
the Quantikine kit standards. These results indicate that this kit can
be used to determine relative mass values for natural human Amphiregulin.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Recombinant and natural human Amphiregulin
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, EDTA Plasma, Heparin Plasma, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 102 | 314 | 631 | 99.4 | 308 | 614 |
| Standard Deviation | 5.63 | 10.5 | 22.4 | 9.85 | 20.5 | 28.8 |
| CV% | 5.5 | 3.3 | 3.5 | 9.9 | 6.7 | 4.7 |
Recovery for Human Amphiregulin Quantikine ELISA Kit
The recovery of Amphiregulin spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 95 | 88-101 |
| EDTA Plasma (n=4) | 93 | 82-105 |
| Heparin Plasma (n=4) | 94 | 82-103 |
| Saliva (n=4) | 104 | 98-113 |
| Serum (n=4) | 95 | 85-103 |
| Urine (n=4) | 88 | 75-99 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Amphiregulin were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Amphiregulin Quantikine ELISA Kit
Human Amphiregulin ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Amphiregulin
Additional Amphiregulin Products
Product Documents for Human Amphiregulin Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Amphiregulin Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Amphiregulin Quantikine ELISA Kit
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Customer Reviews for Human Amphiregulin Quantikine ELISA Kit (6)
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6 Customer Ratings
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Sample Tested: Prostate cancerVerified Customer | Posted 11/14/2020
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Sample Tested: colorectal adenocarcinoma cell lineVerified Customer | Posted 04/18/2018
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Sample Tested: human pancreatic cell lineVerified Customer | Posted 04/05/2018
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Sample Tested: SerumVerified Customer | Posted 01/11/2018
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Sample Tested: SUM-159PT human breast cancer cell lineVerified Customer | Posted 10/20/2017Very high sensitivity - no problem detecting amphiregulin released from human breast cancer cells.
-
Sample Tested: EDTA PlasmaVerified Customer | Posted 09/07/2017The Amphiregulin Quantikine ELISA kit performed as expected with low % CV's, and reproducible results.
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Protocols
View specific protocols for Human Amphiregulin Quantikine ELISA Kit (DAR00):
Refer to the product for complete assay procedure.
The conjugate must remain at 2-8 °C during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours on the benchtop.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL cold Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Human Amphiregulin Quantikine ELISA Kit
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Q: Assay Diluent RD1-14 contains visible precipitate. Is this acceptable to use?
A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-14. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.
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