A part of MilliporeSigma

17-460 | EGFR STAR ELISA Kit

96 assays  
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      Replacement Information

      Key Specifications Table

      Analytes AvailableSpecies ReactivityKey ApplicationsDetection Methods
      EGFR H ELISA Chromogenic
      Catalogue Number17-460
      Brand Family Upstate
      Trade Name
      • STAR
      • Upstate
      DescriptionEGFR STAR ELISA Kit
      Alternate Names
      • Epidermal growth factor receptor
      • Proto-oncogene c-ErbB-1
      • Receptor tyrosine-protein kinase erbB-1
      • EGF Receptor
      • EGFR
      • ERBB
      • ERBB1
      Background InformationI. TEST PRINCIPLE
      The UPSTATE® colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The EGFR plate is coated with a specific mouse monoclonal anti-EGFR capture antibody. Sample lysate or the standard included in the kit are incubated in the microwells allowing EGFR antigen to be captured in the plate wells. The plate is then washed to remove any un-bound non-nspecific material. A specific rabbit anti-EGFR antibody is added to detect the captured full length EGFR on the plate well. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader. This allows for a sensitive enzymatic detection of the sample.
      The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to to generate a standard curve for EGFR measurement.

      Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that plays a key role in the regulation of essential normal cellular processes and in the pathophysiology of hyperproliferative diseases such as cancer. Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase known to be essential for mediation of both proliferative and survival signals to cells. Activation of the EGFR signaling pathway has been linked with increased cell proliferation, angiogenesis, metastasis and decreased apoptosis. EGFR is found in most solid tumors and its autophosphorylation activates the intrinsic kinase activity toward heterologous substrates, as well as creating docking sites for adapter proteins for multiple adapter proteins to bind and nucleate signaling complexes that activate the Ras, PI3 Kinase, and PLC pathways.
      Many of the EGFR mutations are directly related to various diseases and are the target of numerous drugs. For instance, the amino acid substitution L858R is one of several heterozygous mutations that have been identified in Non-Small-Cell Lung Cancer (NSCLC) patients who have clinical responses to the EGFR inhibitor Iressa®. There is some evidence that these mutations result in elevated activity and enhanced sensitivity to Iressa®. In patients with tumors bearing Iressa®-sensitive mutations, resistant subclones containing an additional EGFR mutation, T790M, emerge in the presence of the drug. It has been shown experimentally that the T790M mutation leads to high-level functional resistance to Iressa. In patients with tumors bearing Iressa®-sensitive mutations (eg. L858R, L861Q), resistant subclones containing the T790M mutation emerge in the presence of the drug.
      Materials Required but Not Delivered1. Multi-channel or repeating pipettes
      2. Plate shaker (optional)
      3. Pipettors and tips capable of accurately measuring 1-1000 µL
      4. Graduated serological pipettes
      5. 96-well microtiter Plate Reader with 450 nm filter
      6. Graphing software for plotting data or graph paper for manual plotting of data
      7. Microfuge tubes for standard and sample dilutions
      8. Mechanical vortex
      9. 1 liter container
      10. Distilled or deionized water
      Product Information
      • Capture Plate pre-coated with anti-EGFR antibody: (Part No. 17-460A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.
      • Anti-EGFR detection antibody: (Part No. 17-460B) One bottle (11 mL) of anti-EGFR detection antibody containing sodium azide, ready to use.
      • ELISA Diluent: (Part No. 17-460C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.
      • 25X ELISA Wash Buffer: (Part No. 17-460D) One bottle (50 mL) of 25X ELISA Wash Buffer.
      • Anti-Rabbit IgG HRP conjugate: (Part No. 17-460E) One vial (125 µL) of 100X anti-rabbit HRP conjugate containing thimerosal.
      • HRP Diluent: (Part No. 17-460F) One bottle (25 mL) of HRP Diluent containing thimerosal.
      • TMB Solution: (Part No. 17-460G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use.
      • Stop Solution: (Part No. 17-460H) One bottle (25 mL) of stop solution, ready to use.
      • EGFR Standard: (Part No. 17-460I) Two vials of EGFR (full length) standard, lyophilized.
      • Plate Covers: Two plate covers.
      Detection methodChromogenic
      Key Applications
      • ELISA
      Biological Information
      Species Reactivity
      • Human
      Analytes Available
      • EGFR
      Entrez Gene Number
      Gene Symbol
      • EGFR
      • ERBB1
      • ERBB
      • mENA
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P00533 # Isoform 2/truncated isoform may act as an antagonist.
      SIZE: 1210 amino acids; 134277 Da
      SUBUNIT: Binds RIPK1. CBL interacts with the autophosphorylated C- terminal tail of the EGF receptor. Part of a complex with ERBB2 and either PIK3C2A or PIK3C2B. The autophosphorylated form interacts with PIK3C2B, maybe indirectly. Interacts with PELP1.
      SUBCELLULAR LOCATION: Cell membrane; Single-pass type I membrane protein. & Isoform 2: Secreted.
      TISSUE SPECIFICITY: Expressed in placenta. Isoform 2 is also expressed in ovarian cancers.
      PTM: Phosphorylation of Ser-695 is partial and occurs only if Thr- 693 is phosphorylated. & Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occur.DISEASE:SwissProt: P00533 # Defects in EGFR are associated with lung cancer [MIM:211980].
      SIMILARITY: SwissProt: P00533 ## Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily. & Contains 1 protein kinase domain.
      MISCELLANEOUS: Binding of EGF to the receptor leads to dimerization, internalization of the EGF-receptor complex, induction of the tyrosine kinase activity, stimulation of cell DNA synthesis, and cell proliferation.
      Physicochemical Information
      • Sensitivity: 0.1 ng/mL
        Range of Detection: 0.16 to 10 ng/mL
        Species Reactivity: Human
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by ELISA.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain the unopened kit at 2-8°C until expiration date.
      Packaging Information
      Material Size96 assays
      Transport Information
      Supplemental Information


      Certificates of Analysis

      TitleLot Number
      STAR EGFR ELISA Kit - Manual Manual


      Reference overviewPub Med ID
      Functional characterization of gefitinib uptake in non-small cell lung cancer cell lines.
      Galetti, Maricla, et al.
      Biochem. Pharmacol., 80: 179-87 (2010) 2010

      Show Abstract
      20363215 20363215

      Technical Info

      96 well Template 2 Up
      96 well Template Single


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      User Guides

      Instruction Manual For STAR EGFR ELISA Kit

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