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AB284728

Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free

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Rabbit Monoclonal EGFR phospho T669 antibody. Carrier free. Suitable for Dot, WB, ICC/IF and reacts with Synthetic peptide, Human samples.

View Alternative Names

ERBB, ERBB1, HER1, EGFR, Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1

3 Images
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)

This data was developed using ab75980, the same antibody clone in a different buffer.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 cells labelling EGFR with ab75980 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/1000 dilution. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counterstain was DAPI (Blue).

Western blot - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)
  • WB

Supplier Data

Western blot - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)

This data was developed using ab75980, the same antibody clone in a different buffer.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-EGFR (phospho T669) antibody [EP2256Y] (<a href='/en-us/products/primary-antibodies/egfr-phospho-t669-antibody-ep2256y-ab75980'>ab75980</a>) at 1/2000 dilution

Lane 1:

Untreated A431 (Human epidermoid carcinoma) whole cell lysate at 15 µg

Lane 2:

A431 treated with 100ng/ml Epidermal Growth Factor (EGF) for 30mins whole cell lysate at 15 µg

Lane 3:

A431 treated with 100ng/ml Epidermal Growth Factor (EGF) for 30mins whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 170 kDa

false

Exposure time: 40s

Dot Blot - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)
  • Dot

Supplier Data

Dot Blot - Anti-EGFR (phospho T669) antibody [EP2256Y] - BSA and Azide free (AB284728)

This data was developed using ab75980, the same antibody clone in a different buffer.

Dot blot analysis of EGFR (phospho T669) using ab75980 at 1/1000 dilution, followed by a Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated (ab97051) at 1/2500 dilution. Blocking and dilution buffer : 5% NFDM/TBST. Exposure time : 41s.

All lanes:

Dot Blot - Anti-EGFR (phospho T669) antibody [EP2256Y] (<a href='/en-us/products/primary-antibodies/egfr-phospho-t669-antibody-ep2256y-ab75980'>ab75980</a>) at 1/1000 dilution

Lane 1:

EGFR (phospho T669) peptide

Lane 2:

EGFR non-phospho peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2500 dilution

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2256Y

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

ICC/IF, WB, Dot

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Recognises EGFR phosphorylated on Threonine 669 of the mature human isoform 1 (corresponding to T693 from the precursor form P00533-1/p170)

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Reactivity data

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Product details

ab284728 is the carrier-free version of ab75980.

This product has switched from a hybridoma to recombinant production method on 9th February 2024.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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Supplementary information

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

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.

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.
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Product protocols

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

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.
See full target information EGFR phospho T669
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Product promise

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For full details, please see our Terms & Conditions

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