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Human EGFR (Tyr1068) In-Cell ELISA Kit is Cell-based (qualitative) ELISA for the measurement of Human EGFR (Tyr1068) production in Human in Adherent cells samples.

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Images

In-Cell ELISA - Human EGFR (Tyr1068) In-Cell ELISA Kit (AB126420), expandable thumbnail
  • Western blot - Human EGFR (Tyr1068) In-Cell ELISA Kit (AB126420), expandable thumbnail

Key facts

Detection method
Colorimetric
Sample types
Adherent cells
Assay type
Cell-based (qualitative)
Reactive species
Human
Assay time
5h 10m

Reactivity data

Application
ELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

The protein expressed by the EGFR gene functions as a receptor tyrosine kinase, binding with ligands from the EGF family to activate several signaling cascades that translate extracellular signals into cellular responses. Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen, BTC/betacellulin, epiregulin, and HBEGF. Ligand binding triggers receptor dimerization and autophosphorylation, recruiting adapter proteins like GRB2 and activating complex downstream signaling pathways such as RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC, and STATs, and possibly the NF-kappa-B cascade. EGFR also directly phosphorylates proteins like RGS16 and MUC1, influencing various cellular processes including its coupling with G protein-coupled receptor signaling and cell migration through interaction with CCDC88A/GIV. Isoform 2 of EGFR may act as an antagonist to EGF actions. In microbial infection, EGFR also serves as a receptor facilitating hepatitis C virus (HCV) entry into hepatocytes by promoting CD81-CLDN1 receptor complexes and enhancing membrane fusion with HCV envelope glycoproteins. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

What's included?

1 x 96 Tests
Components
Anti-Rabbit IgG Concentrate (Item I-1)
1 x 10 µL
Blocking Buffer Concentrate (5X)
1 x 20 mL
Fixing Solution
1 x 30 mL
Quenching Buffer Concentrate (30x)
1 x 2 mL
Rabbit Anti-EGFR Concentrate (Item H)
1 x 6 µL
Rabbit Anti-Phospho-EGFR (Tyr1068) Concentrate (Item G)
1 x 6 µL
Stop Solution
1 x 14 mL
TMB One-Step Substrate Reagent
1 x 12 mL
Uncoated 96-well Microplate
1 x 1 Unit
Wash Buffer A Concentrate (20X)
1 x 30 mL
Wash Buffer B Concentrate (20X)
1 x 30 mL

Recommended products

Human EGFR (Tyr1068) In-Cell ELISA Kit is Cell-based (qualitative) ELISA for the measurement of Human EGFR (Tyr1068) production in Human in Adherent cells samples.

Key facts

Detection method
Colorimetric
Sample types
Adherent cells
Assay type
Cell-based (qualitative)
Reactive species
Human
Assay time
5h 10m
Assay Platform
Microplate

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

ab126420 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 (Tyr1068) 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 (Tyr1068) 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 (Tyr1068) or anti-EGFR (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-rabbit 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

EGFR or Epidermal Growth Factor Receptor is a transmembrane glycoprotein that acts as a receptor for members of the epidermal growth factor family. Known alternatively as ErbB1 or HER1 this receptor has an approximate molecular weight of 170 kDa. EGFR is expressed in various cell types notably on epithelial cells and can influence multiple cellular processes through its kinase activity. It participates in the regulation of cell growth multiplication and survival by activating its kinase domain upon ligand binding.

Biological function summary

The EGFR protein plays an important role in cellular communication and signaling processes. EGFR pairs with other receptor family members to form active dimers or even higher-order complexes which in turn initiate intracellular signaling cascades. Through these complexes EGFR influences many processes including cell differentiation and repair. This function of EGFR makes it an integral part of mammalian biology affecting how cells respond to their environment by mediating changes in gene expression.

Pathways

EGFR is a central player in the MAPK and PI3K/Akt signaling pathways. Alongside other protein partners like KRAS and PI3 kinase it contributes to transmitting signals from the cell surface to the nucleus affecting gene transcription and cell behavior. These pathways are important for normal cell growth and division and aberrations in these pathways can lead to excessive or insufficient cell proliferation.

Associated diseases and disorders

EGFR is pertinent to cancer biology including non-small cell lung cancer and glioblastoma where mutations or overexpression of the receptor frequently occur. It connects to proteins such as PTEN and BRAF which influence tumor progression and response to targeted therapies. EGFR's involvement in these disorders highlights its significance as a therapeutic target since it can be manipulated to alter disease progression.

Product promise

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2 product images

  • In-Cell ELISA - Human EGFR (Tyr1068) In-Cell ELISA Kit (ab126420), expandable thumbnail

    In-Cell ELISA - Human EGFR (Tyr1068) In-Cell ELISA Kit (ab126420)

    A431 cells were stimulated by different concentrations of EGF for 10 min at 37°C.

  • Western blot - Human EGFR (Tyr1068) In-Cell ELISA Kit (ab126420), expandable thumbnail

    Western blot - Human EGFR (Tyr1068) In-Cell ELISA Kit (ab126420)

    Western blot analysis of extracts from 100 ng/ml hEGF treated A431 cells. Phospho-EGFR (Tyr1068) and EGFR antibodies were used in both detection assays.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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