ELISA Kit for Epidermal Growth Factor Receptor (EGFR) Rattus norvegicus (Rat) Sandwich ELISA

ErbB-1; ErbB1; ER1; ERB-B1; MENA; HER1; Erythroblastic Leukemia Viral(v-Erb-B)oncogene Homolog,Avian; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1

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  • ELISA Kit for Epidermal Growth Factor Receptor (EGFR) Packages (Simulation)
  • ELISA Kit for Epidermal Growth Factor Receptor (EGFR) Packages (Simulation)
  • ELISA Kit for Epidermal Growth Factor Receptor (EGFR) Results demonstration
  • SEA757Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Epidermal Growth Factor Receptor (EGFR) and the recovery rates were calculated by comparing the measured value to the expected amount of Epidermal Growth Factor Receptor (EGFR) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-102 81
EDTA plasma(n=5) 80-102 87
heparin plasma(n=5) 90-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Epidermal Growth Factor Receptor (EGFR) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Epidermal Growth Factor Receptor (EGFR) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Epidermal Growth Factor Receptor (EGFR) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-101% 93-102% 82-99% 84-103%
EDTA plasma(n=5) 91-101% 78-89% 85-99% 78-98%
heparin plasma(n=5) 97-105% 95-102% 83-92% 78-93%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Epidermal Growth Factor Receptor (EGFR)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Epidermal Growth Factor Receptor (EGFR). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Epidermal Growth Factor Receptor (EGFR). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Epidermal Growth Factor Receptor (EGFR), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Epidermal Growth Factor Receptor (EGFR) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Nimotuzumab combined with gemcitabine and cisplatin as second-line chemotherapy for advanced non-small-cell lung cancerWiley: source
  • Increased Epidermal Growth Factor Receptor (EGFR) Associated with Hepatocyte Growth Factor (HGF) and Symptom Severity in Children with Autism …Pubmed:25249767
  • GC1118, an anti-EGFR antibody with a distinct binding epitope and superior inhibitory activity against high-affinity EGFR ligandsPubMed: 26586721
  • LINC01225 promotes occurrence and metastasis of hepatocellular carcinoma in an epidermal growth factor receptor-dependent pathwayPubmed:26938303
  • Microfluidic immunosensor based on mesoporous silica platform and CMK-3/poly-acrylamide-co-methacrylate of dihydrolipoic acid modified gold electrode for cancer biomarker detection.pubmed:28335979
  • EGFR detection in extracellular vesicles of breast cancer patients through immunosensor based on silica-chitosan nanoplatformDoi: 10.1016/j.talanta.2018.10.016
  • Design, synthesis, molecular docking and biological activity evaluation of some novel indole derivatives as potent anticancer active agents and apoptosis inducersPubmed: 30665034
  • Novel thienopyrimidine derivatives as dual EGFR and VEGFR-2 inhibitors: design, synthesis, anticancer activity and effect on cell cycle profilePubmed: 30919701
  • Design and synthesis of new benzoxazole/benzothiazole-phthalimide hybrids as antitumor-apoptotic agents
  • Synthesis and anticancer activity of bis-benzo [d][1, 3] dioxol-5-yl thiourea derivatives with molecular docking studyPubmed: 31288134
  • Evaluation of the Effect Derived from Silybin with Vitamin D and Vitamin E Administration on Clinical, Metabolic, Endothelial Dysfunction, Oxidative Stress Parameters …Pubmed: 31737175
  • Patient-derived xenograft (PDX) models of colorectal carcinoma (CRC) as a platform for chemosensitivity and biomarker analysis in personalized medicinePubmed: 33212364
  • Association between Serum Epidermal Growth Factor Receptor and Cyclooxygenase-2 Levels in Patients with Non-small Cell Carcinoma of Lung
  • Synthesis, EGFR-TK inhibition and anticancer activity of new quinoxaline derivatives
  • Humoral immune response to epidermal growth factor receptor in lung cancer33495907
  • Pyrazolo[3,4-d]pyrimidine-based dual EGFR T790M/HER2 inhibitors: Design, synthesis, structure¨Cactivity relationship and biological activity as potential antitumor and anticonvulsant agents33545637
  • Design, synthesis, antiproliferative evaluation, and molecular docking study of new quinoxaline derivatives as apoptotic inducers and EGFR inhibitors
  • Emitter–Quencher Pair of Single Atomic Co Sites and Monolayer Titanium Carbide MXenes for Luminol Chemiluminescent Reactions34914377
  • Discovery of new pyrimido [5, 4-c] quinolines as potential antiproliferative agents with multitarget actions: Rapid synthesis, docking, and ADME studiesPubmed:35219045

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