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AB319114

Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free

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Rabbit Recombinant Multiclonal EGFR phospho Y845 + Y1068 + Y1086 antibody. Carrier free. Suitable for Dot, ICC/IF, IP, WB and reacts with Synthetic peptide - Human, Human samples.

View Alternative Names

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

5 Images
Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)

This data was developed using ab319113, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling EGFR (phospho Y845 + Y1068 + Y1086) with ab319113 at 1/10000 (0.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing increased membranous staining in A431 cells treated with EGF (100 ng/ml) for 30 min (shown in green). The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

Immunoprecipitation - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)
  • IP

Supplier Data

Immunoprecipitation - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)

This data was developed using ab319113, the same antibody clone in a different buffer formulation.

EGFR (phospho Y845 + Y1068 + Y1086) was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate with ab319113 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab319113 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate 2 ug
Lane 2 : ab319113 IP in A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab319113 in A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (<a href='/en-us/products/primary-antibodies/egfr-phospho-y845-y1068-y1086-antibody-rm1132-ab319113'>ab319113</a>) at 1/30 dilution

All lanes:

A431 (human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30min whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)
  • WB

Supplier Data

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)

This data was developed using ab319113, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-EGFR antibody [EP38Y] (ab52894) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (<a href='/en-us/products/primary-antibodies/egfr-phospho-y845-y1068-y1086-antibody-rm1132-ab319113'>ab319113</a>) at 1/5000 dilution

Lane 1:

Untreated A431 (human epidermoid carcinoma epithelial cell) whole cell lysate (untreated membrane) at 10 µg

Lane 2:

A431 treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 10 µg

Lane 3:

Untreated A431 whole cell lysate (alkaline phosphatase treated membrane) at 10 µg

Lane 4:

A431 treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 10 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 1/100000 dilution

Observed band size: 175 kDa,124 kDa

false

Exposure time: 169s

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)
  • WB

Supplier Data

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)

This data was developed using ab319113, the same antibody clone in a different buffer formulation.

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

In Western blot, ab319113 was shown to bind specifically to EGFR. Target of interest was observed at 175kDa, in wild-type HeLa cell lysates (lane 1/2/5/6) with no signal observed at this size in EGFR knockout cell line (lane 3/4/7/8) (knockout cell line ab255385 / knockout cell lysate ab263845).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-EGFR antibody [EP38Y] (ab52894) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (<a href='/en-us/products/primary-antibodies/egfr-phospho-y845-y1068-y1086-antibody-rm1132-ab319113'>ab319113</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Wild-type HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated EGFR knockout HeLa whole cell lysate (untreated membrane) at 20 µg

Lane 4:

EGFR knockout HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 5:

Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 6:

Wild-type HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 7:

Untreated EGFR knockout HeLa whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 8:

EGFR knockout HeLa treated with 100ng/ml EGF for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 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 1/100000 dilution

Observed band size: 175 kDa,36 kDa

false

Exposure time: 180s

Dot Blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)
  • Dot

Supplier Data

Dot Blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] - BSA and Azide free (AB319114)

This data was developed using ab319113, the same antibody clone in a different buffer formulation.

Dot blot analysis of EGFR (phospho Y845 + Y1068 + Y1086) using ab319113 at 1 : 1000 (0.496 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human EGFR (phospho Y1068) peptide
Lane2 : Human EGFR non-phospho peptide
Lane3 : Human EGFR (phospho Y1086) peptide
Lane4 : Human EGFR non-phospho peptide
Lane5 : Human EGFR (phospho Y845) peptide
Lane6 : Human EGFR non-phospho peptide

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

All lanes:

Dot Blot - Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132] (<a href='/en-us/products/primary-antibodies/egfr-phospho-y845-y1068-y1086-antibody-rm1132-ab319113'>ab319113</a>) at 1/1000 dilution

Lane 1:

Human EGFR (phospho Y1068) peptide

Lanes 2, 4 and 6:

Human EGFR non-phospho peptide

Lane 3:

Human EGFR (phospho Y1086) peptide

Lane 5:

Human EGFR (phospho Y845) 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 1/100000 dilution

false

Exposure time: 180s

  • Unconjugated

    Anti-EGFR (phospho Y845 + Y1068 + Y1086) antibody [RM1132]

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1132

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IP, WB, ICC/IF, Dot

applications

Immunogen

This product was produced with the following immunogens:

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

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>" }, "Mouse": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "" }, "Rat": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "" }, "Synthetic peptide - Human": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "" } } }
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Product details

ab319114 is the carrier-free version of ab319113.

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

  • - The sensitivity of polyclonal antibodies by recognising multiple epitopes
  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

View our range of recombinant multiclonal antibodies.

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.

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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

Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses (PubMed : 10805725, PubMed : 27153536, PubMed : 2790960, PubMed : 35538033). Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF (PubMed : 12297049, PubMed : 15611079, PubMed : 17909029, PubMed : 20837704, PubMed : 27153536, PubMed : 2790960, PubMed : 7679104, PubMed : 8144591, PubMed : 9419975). Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules (PubMed : 27153536). May also activate the NF-kappa-B signaling cascade (PubMed : 11116146). Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling (PubMed : 11602604). Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin (PubMed : 11483589). Positively regulates cell migration via interaction with CCDC88A/GIV which retains EGFR at the cell membrane following ligand stimulation, promoting EGFR signaling which triggers cell migration (PubMed : 20462955). Plays a role in enhancing learning and memory performance (By similarity). Plays a role in mammalian pain signaling (long-lasting hypersensitivity) (By similarity).. Isoform 2 may act as an antagonist of EGF action.. (Microbial infection) Acts as a receptor for hepatitis C virus (HCV) in hepatocytes and facilitates its cell entry. Mediates HCV entry by promoting the formation of the CD81-CLDN1 receptor complexes that are essential for HCV entry and by enhancing membrane fusion of cells expressing HCV envelope glycoproteins.
See full target information EGFR phospho Y845 + Y1068 + Y1086
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Product promise

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For full details, please see our Terms & Conditions

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