EGFR ELISA Kit (Human) (OKBB00137)

Catalog No: OKBB00137

Size:96 Tests

Human EGFR ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human EGFR ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for EGFR has been precoated onto 96-well plates. Standards(NS0, M1-S645) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for EGFR is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human EGFR amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	1.37	4.68	6.21	1.49	4.92	7.14
  St. Dev.	0.053	0.211	0.329	0.073	0.32	0.614
  %CV	3.86	4.50	5.29	4.89	6.50	8.59

• ELISA Kit Component:
  • Lyophilized recombinant human EGFR standard: 10 ng/tube x 2
  • Anti-human EGFR Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human EGFR antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s human EGFR ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human EGFR specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human EGFR amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands.1 It is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR exists on the cell surface and is activated by binding of its specific ligands, including epidermal growth factor and transforming growth factor α (TGFα). EGFR and its ligands are cell signaling molecules involved in diverse cellular functions, including cell proliferation, differentiation, motility, and survival, and in tissue development.2 Mutations that lead to EGFR overexpression (known as upregulation) or overactivity have been associated with a number of cancers, including lung cancer and glioblastoma multiforme. In this latter case a more or less specific mutation of EGFR, called EGFRvIII is often observed.3
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum, plasma (heparin, EDTA) and human milk
• Sensitivity: <1 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Herbst RS (2004). "Review of epidermal growth factor receptor biology". Int. J. Radiat. Oncol. Biol. Phys. 59 (2
  Suppl): 21–6.
  2. Wang, K.; Yamamoto, H.; Chin, J. R.; Werb, Z.; Vu, T. H.: Epidermal growth factor receptor-deficient mice have
  delayed primary endochondral ossification because of defective osteoclast recruitment. J. Biol. Chem. 279:
  53848-53856, 2004.
  3. Kuan CT, Wikstrand CJ, Bigner DD (June 2001). "EGF mutant receptor vIII as a molecular target in cancer therapy".
  Endocr. Relat. Cancer 8 (2): 83–96.
• Specificity: Natural and recombinant human EGFR
• Immunogen: Expression system for standard: NS0; Immunogen sequence: M1-S645
• Description: For quantitative detection of human EGFR in cell culture supernatants, serum, plasma(heparin, EDTA) and human milk.

Target Info

• Gene Symbol: EGFR
• Gene Full Name: epidermal growth factor receptor
• Alias Symbols: 9030024J15Rik, AI552599, Avian erythroblastic leukemia viral (v erb b) oncogene homolog, Avian erythroblastic leukemia viral (verbb) oncogene homolog, Cell growth inhibiting protein 40, Cell proliferation inducing protein 61, EGF R, EGFR, EGFR_HUMAN, Epidermal growth factor receptor, Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog), Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian)Erbb, erbb 1, Erbb1, Erythroblastic leukemia viral (v erb-b) oncogene homolog, HER1, mENA, Oncogene ERBB, OTTMUSP00000005385, OTTMUSP00000005386, PIG61, Proto oncogene c ErbB 1, Proto-oncogene c-ErbB-1, Receptor tyrosine protein kinase ErbB 1, Receptor tyrosine protein kinase ErbB1, Receptor tyrosine-protein kinase ErbB-1, RP23-295E4.1, Urogastrone, wa2, Wa5
• NCBI Gene Id: 1956
• Protein Name: Epidermal growth factor receptor
• Description of Target: The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands.1 It is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR exists on the cell surface and is activated by binding of its specific ligands, including epidermal growth factor and transforming growth factor α (TGFα). EGFR and its ligands are cell signaling molecules involved in diverse cellular functions, including cell proliferation, differentiation, motility, and survival, and in tissue development.2 Mutations that lead to EGFR overexpression (known as upregulation) or overactivity have been associated with a number of cancers, including lung cancer and glioblastoma multiforme. In this latter case a more or less specific mutation of EGFR, called EGFRvIII is often observed.3
• Uniprot ID: P00533
• Protein Accession #: NP_005219.2
• Nucleotide Accession #: NM_005228.3