Human EGFR Quantikine ELISA Kit

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DEGFR0
Control Products Available
EGFR ELISA Kit ELISA
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Human EGFR Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Human Milk (50 uL)
Sensitivity
0.036 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Human Milk)
Specificity
Natural and recombinant human EGF R
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.

Product Summary

The Quantikine Human EGF R immunoassay is a 4.5 hour solid phase ELISA designed to measure EGF R in cell culture supernates, serum, plasma, and human milk. It contains NS0-expressed recombinant human EGF R and has been shown to accurately quantitate the recombinant factor. Results obtained using natural EGF R 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 naturally occurring human EGF R.

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, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1.65 5.08 10.6 1.63 5 10
Standard Deviation 0.094 0.17 0.507 0.144 0.416 0.816
CV% 5.7 3.3 4.8 8.8 8.3 8.2

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1.69 5.33 11.2 1.7 5.21 10.4
Standard Deviation 0.072 0.199 0.616 0.161 0.522 1.03
CV% 4.3 3.7 5.5 9.4 10 10

Recovery

The recovery of EGF R spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 86-106
EDTA Plasma (n=4) 99 91-108
Heparin Plasma (n=4) 101 92-111
Serum (n=4) 101 89-117

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of EGF R were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human EGF R/ErbB1 ELISA Linearity

Data Examples

Human EGF R/ErbB1 ELISA Cell Culture Supernate/Human Milk Standard Curve

Human EGF R/ErbB1 ELISA Serum/Plasma Standard Curve

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: EGFR

The EGF R subfamily of receptor tyrosine kinases comprises four members: EGF R (also known as HER-1, ErbB1, or ErbB), ErbB2 (Neu, HER-2), ErbB3 (HER-3), and ErbB4 (HER-4). All family members are type I transmembrane glycoproteins with an extracellular ligand binding domain containing two cysteine-rich domains separated by a spacer region and a cytoplasmic domain containing a membrane-proximal tyrosine kinase domain followed by multiple tyrosine autophosphorylation sites. The human EGF R cDNA encodes a 1210 amino acid (aa) precursor with a 24 aa signal peptide, a 621 aa extracellular domain (ECD), a 23 aa transmembrane segment, and a 542 aa cytoplasmic domain. Soluble receptors consisting of the extracellular ligand binding domain are generated by alternate splicing in human and mouse. Within the ECD, human EGF R shares 88% aa sequence identity with mouse and rat EGF R. It shares 43% - 44% aa sequence identity with the ECD of human ErbB2, ErbB3, and ErbB4. EGF R binds a subset of the EGF family ligands, including EGF, amphiregulin, TGF-alpha, betacellulin, epiregulin, HB-EGF, and epigen. Ligand binding induces EGF R homodimerization as well as heterodimerization with ErbB2, resulting in kinase activation, heterodimerization tyrosine phosphorylation and cell signaling. EGF R can also be recruited to form heterodimers with the ligand-activated ErbB3 or ErbB4. EGF R signaling regulates multiple biological functions including cell proliferation, differentiation, motility, and apoptosis. EGF R is overexpressed in a wide variety of tumors and is the target of several anti-cancer drugs.

Long Name:
Epidermal Growth Factor Receptor
Entrez Gene IDs:
1956 (Human); 13649 (Mouse)
Alternate Names:
avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; EC 2.7.10; EC 2.7.10.1; EGF R; EGFR; epidermal growth factor receptor (avian erythroblastic leukemia viral (v-erb-b)oncogene homolog); epidermal growth factor receptor; ErbB; ErbB1; ERBB1PIG61; HER1; HER-1; mENA; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1
⚠ 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.

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

Citations for Human EGFR Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Interleukin 4, interleukin 6 and osteopontin-serological markers of head and neck malignancy in primary diagnostics: A pilot study.
    Authors: Aderhold C, Grobschmidt G, Sauter A, Faber A, Hormann K, Schultz J
    Oncol Lett, 2014;8(3):1112-1118.
    Species: Human
    Sample Types: Serum
  2. Cancer cells induced to express mesenchymal phenotype release exosome-like extracellular vesicles carrying tissue factor.
    Authors: Garnier, Delphine, Magnus, Nathalie, Lee, Tae Hoon, Bentley, Victoria, Meehan, Brian, Milsom, Chloe, Montermini, Laura, Kislinger, Thomas, Rak, Janusz
    J Biol Chem, 2012;287(52):43565-72.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Morphological and biochemical features of obesity are associated with mineralization genes' polymorphisms.
    Authors: Korostishevsky M, Cohen Z, Malkin I
    Int J Obes (Lond), 2010;34(8):1308-18.
    Species: Human
    Sample Types: Plasma

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