人EGFR (pY992) + total EGFR ELISA试剂盒(ab126443)

概述

  • 产品名称
    人EGFR (pY992) + total EGFR ELISA试剂盒
  • 检测方法
    Colorimetric
  • 样品类型
    Cell Lysate
  • 检测类型
    Semi-quantitative
  • 检测时间
    5h 00m
  • 实验步骤
    Multiple steps standard assay
  • 种属反应性
    与反应: Human
  • 产品概述

    ab126443 kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phosphorylated EGFR protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-EGFR (Tyr 992) and total EGFR (help normalize the results of phospho-EGFR from different cell lysate being compared). An anti-EGFR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-phosphorylated EGFR (Tyr 992) or anti-total-EGFR antibody is used to detect phosphorylated or non-phosphorylated EGFR. After washing away unbound antibody, HRP-conjugated anti-Rabbit IgG or HRP-Streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of EGFR (Tyr992) or total EGFR bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • 经测试应用
    适用于: Sandwich ELISAmore details
  • 平台
    Microplate

性能

  • 存放说明
    Store at -20°C. Please refer to protocols.
  • 组件 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2x Cell lysate buffer 1 x 5ml
    5X Assay Diluent 1 x 15ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Anti-phospho-EGFR (Tyr 992) 1 vial
    Biotinylated-Anti-EGFR 1 vial
    EGFR Microplate (12 strips x 8 wells) coated with monoclonal anti-EGFR 1 unit
    HRP-conjugated anti-rabbit IgG 1 x 25µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • 研究领域
  • 功能
    Receptor for EGF, but also for other members of the EGF family, as TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation. Phosphorylates MUC1 in breast cancer cells and increases the interaction of MUC1 with SRC and CTNNB1/beta-catenin.
    Isoform 2 may act as an antagonist of EGF action.
  • 组织特异性
    Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
  • 疾病相关
    Defects in EGFR are associated with lung cancer (LNCR) [MIM:211980].
  • 序列相似性
    Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰
    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.
  • 细胞定位
    Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Co-localizes with TNK2 on the endosomes.
  • Information by UniProt
  • 别名
    • Avian erythroblastic leukemia viral (v erb b) oncogene homolog
    • Cell growth inhibiting protein 40
    • Cell proliferation inducing protein 61
    • EGFR
    • EGFR_HUMAN
    • Epidermal growth factor receptor
    • ERBB
    • ERBB1
    • HER1
    • mENA
    • Oncogene ERBB
    • PIG61
    • Proto-oncogene c-ErbB-1
    • Receptor tyrosine-protein kinase ErbB-1
    • v ERB B avian erythroblastic leukemia viral oncogene homolog
    see all
  • 数据库链接

应用

Our Abpromise guarantee covers the use of ab126443 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Sandwich ELISA Use at an assay dependent concentration.

图片

  • A431 cells were treated with recombinant human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed using ab126443.
  • A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 10 min. Cell lysates were analyzed by Western Blot.
  • The A431 cells were treated with 100 ng/mL recombinant human EGF for 20 minutes to induce phosphorylation of EGFR. Serial dilutions of lysates were analyzed using ab126443.

实验方案

文献

ab126443 has not yet been referenced specifically in any publications.

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