人EGFR In-Cell ELISA试剂盒(ab126419)

概述

  • 产品名称
    人EGFR In-Cell ELISA试剂盒
    参阅全部 EGFR 试剂盒
  • 检测方法
    Colorimetric
  • 样品类型
    Adherent cells
  • 检测类型
    Cell-based (qualitative)
  • 检测时间
    5h 10m
  • 实验步骤
    Multiple steps standard assay
  • 种属反应性
    与反应: Human
  • 产品概述

    ab126419 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of EGFR (activated) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured Human cell lines. By determining EGFR protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot.

    In the EGFR Human In-Cell ELISA kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, Anti-Phospho-EGFR (activated) or Anti-EGFR (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti mouse IgG (secondary antibody) is added to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. 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: In-Cell 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
    Anti-Mouse IgG Concentrate (Item I) 1 x 10µl
    5X Blocking Buffer Concentrate (Item F) 1 x 20ml
    Fixing Solution (Item D) 1 x 30ml
    Uncoated 96-well Microplate (Item A) 1 unit
    Mouse Anti-EGFR Concentrate (Item H) 1 x 7µl
    Mouse Anti-Phospho-EGFR (active) Concentrate (Item G) 1 x 7µl
    Quenching Buffer Concentrate (30x) (Item E) 1 x 2ml
    Stop Solution 1 x 14ml
    TMB Substrate Reagent 1 x 12ml
    20X Wash Buffer A Concentrate (Item B) 1 x 30ml
    20X Wash Buffer B Concentrate (Item C) 1 x 30ml
  • 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
    • 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
    see all
  • 数据库链接

应用

Our Abpromise guarantee covers the use of ab126419 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
In-Cell ELISA Use at an assay dependent concentration.

图片

  • A431 cells were stimulated by different concentrations of EGF for 20 min at 37°C.

  • Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-EGFR (activated) and EGFR antibodies were used in both detection assays.
  • A431 cells were stimulated by different concentrations of EGF for 20 min at 37°C.

实验方案

文献

ab126419 has not yet been referenced specifically in any publications.

客户评价及客户问答

The lab let me know the following:

While we cannot say with 100% certainty that this antibody will not cross react with phosphorylated tyrosines on any other molecules besides EGFR, we can predict with pretty high confidence that the antibody ...

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The anti-phospho antibody in the EGFR ELISA kit is a single monoclonal, not a cocktail of several clones. Although it is not available separately, I did find another antibody that may be of interest, Anti-Phosphoserine/threonine/tyrosine, ab15556.
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Thank you for your inquiry.

The lab has confirmed that the phospho EGFR antibody is monoclonal, was purified by protein A chromatography, the concentration is not available (it is not necessary to perform the assay), and the buffer is an aque...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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