EGF (Epidermal Growth Factor) is expressed as a transmembrane protein that is proteolytically cleaved to generate soluble forms. It signals through EGF R/ErbB1 which heterodimerizes with ErbB2, ErbB3, or ErbB4 and can also associate with PDGF R beta and HGF R/c-MET. During development, EGF regulates thymocyte differentiation, neuroglia production, and adipocyte maturation. In the adult, EGF plays a role in mammary gland lactogenesis, fibroblast mitosis, dissociation of the extracellular matrix, and cell migration.
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Human EGF Quantikine ELISA Kit
R&D Systems | Catalog # DEG00
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Key Product Details
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 µL), Serum (10 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Platelet-poor Citrate Plasma (200 µL), Urine (10 µL)
Sensitivity
0.7 pg/mL
Assay Range
3.9-250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Platelet-poor Citrate Plasma, Urine)
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Product Summary for Human EGF Quantikine ELISA Kit
The Quantikine Human EGF Immunoassay kit is a 3.5 or 4.5 hour solid phase ELISA designed to measure EGF levels in cell culture supernates, serum, plasma, and urine. It contains E. coli-derived recombinant human EGF and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human EGF. Results obtained measuring natural human EGF showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural EGF.
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
Natural and recombinant human EGF
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for
the presence of human EGF in this assay. No medical histories were available for the donors
used in this study.
ND=Non-detectable
Cell Culture Supernates - - Human peripheral blood leukocytes (PBL; 1 x 106 cells) were cultured in RPMI 1640 and supplemented with 10% fetal bovine serum. Cells were cultured stimulated with 10 μg/mL PHA for 6 days. An aliquot of the cell culture supernate was removed, assayed for human EGF, and measured 5.4 pg/mL.
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Platelet-poor EDTA plasma (n=37) | 4.87 | 5 | 4.54-5.20 |
| Platelet-poor heparin plasma (n=37) | 4.81 | 16 | ND-5.43 |
| Platelet-poor citrate plasma (n=37) | ND | 0 | _____ |
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=37) | 409 | 106-1063 | 151 |
| Saliva (n=14) | 1703 | 81.8-9600 | 2400 |
| Sample Type | Mean (μg/g Creatinine) | Range (μg/g Creatinine) | Standard Deviation (μg/g Creatinine) |
| Urine (n=11) | 23,019 | 12,352-36,894 | 8524 |
Cell Culture Supernates - - Human peripheral blood leukocytes (PBL; 1 x 106 cells) were cultured in RPMI 1640 and supplemented with 10% fetal bovine serum. Cells were cultured stimulated with 10 μg/mL PHA for 6 days. An aliquot of the cell culture supernate was removed, assayed for human EGF, and measured 5.4 pg/mL.
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.
Cell Culture Supernates, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 15.0 | 78.9 | 134 | 15.0 | 80.0 | 136 |
| Standard Deviation | 0.4 | 2.5 | 4.0 | 0.9 | 4.5 | 6.8 |
| CV% | 2.7 | 3.2 | 3.0 | 6.0 | 5.6 | 5.0 |
EDTA Plasma, Heparin Plasma, Platelet-poor Citrate Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 13.5 | 80 | 133 | 14.7 | 77 | 134 |
| Standard Deviation | 0.6 | 2.0 | 2.7 | 0.9 | 3.3 | 6.1 |
| CV% | 4.4 | 2.5 | 2.0 | 6.1 | 4.3 | 4.6 |
Recovery for Human EGF Quantikine ELISA Kit
The recovery of EGF spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 98 | 88-111 |
| Citrate Plasma (n=10) | 104 | 89-115 |
| EDTA Plasma (n=10) | 100 | 80-111 |
| Heparin Plasma (n=10) | 96 | 79-108 |
| Serum (n=10) | 97 | 80-106 |
| Urine (n=1) | 102 | 97-108 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of EGF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human EGF Quantikine ELISA Kit
Human EGF ELISA Cell Culture Supernate/Urine Standard Curve
Human EGF ELISA Serum/Plasma/Saliva 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: EGF
Long Name
Epidermal Growth Factor
Alternate Names
HOMG4, URG, Urogastrone
Gene Symbol
EGF
Additional EGF Products
Product Documents for Human EGF 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 EGF 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 EGF Quantikine ELISA Kit
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Sample Tested: Urine, Plasma and SerumVerified Customer | Posted 03/15/2022We analyzed urine at a 150-fold dilution and plasma/serum at a 10-fold dilution. The standard curve had an excellent 4PL fit. We continue to use this assay routinely.
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Sample Tested: SerumVerified Customer | Posted 12/11/2021
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Sample Tested: 293T human embryonic kidney cell line and T47D human breast cancer cell lineVerified Customer | Posted 09/13/2018
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Sample Tested: SerumVerified Customer | Posted 06/12/2018
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Sample Tested: Adult thymocytesVerified Customer | Posted 04/03/2018
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Protocols
View specific protocols for Human EGF Quantikine ELISA Kit (DEG00):
Refer to the product for complete assay procedure.
Bring all 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.
- For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.
- Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour. - Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. 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 for Serum & Plasma Samples Only
200 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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