Recombinant Anti-EGFR antibody [EMab-134] - BSA and Azide free (ab264547)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [EMab-134] to EGFR - BSA and Azide free
- Suitable for: WB, Flow Cyt, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-EGFR antibody [EMab-134] - BSA and Azide free
See all EGFR primary antibodies -
Description
Mouse monoclonal [EMab-134] to EGFR - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: WB, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A431 and MDA-MB-468 whole cell lysates. IHC-P: Human oral carcinoma and lung carcinoma tissue; Human skin tissue. Flow Cyt: A431 cells.
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General notes
ab264547 is the carrier-free version of ab264540.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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 production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EMab-134 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab264547 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 130, 180 kDa (predicted molecular weight: 134 kDa).
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Flow Cyt |
1/100.
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IHC-P | (1) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Detects a band of approximately 130, 180 kDa (predicted molecular weight: 134 kDa). |
Flow Cyt
1/100. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. 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. May also activate the NF-kappa-B signaling cascade. 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. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. -
Tissue specificity
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers. -
Involvement in disease
Lung cancer
Inflammatory skin and bowel disease, neonatal, 2 -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.
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 occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197. -
Cellular localization
Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF). - Information by UniProt
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Database links
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
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Alternative names
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
- Cell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibody
see all
Images
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All lanes : Anti-EGFR antibody [EMab-134] (ab264540) at 1/1000 dilution
Lane 1 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
Lane 2 : SW620 (human colorectal adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 134 kDa
Observed band size: 130,180 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1, 2: 15 secs; Lane 3: 8 secs.
The molecular weight observed is in consistent with the literature (PMID: 9090976).
Negative control: SW620 (PMID: 19789347).This image was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (ab264540).
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Immunohistochemical analysis of paraffin-embedded human skin tissue labeling EGFR with ab264540 at 1/100 dilution followed by a ready to use secondary. Membranous staining on human skin is observed. The section was incubated with ab264540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This image was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (ab264540).
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Flow cytometric analysis of SW620 (human colorectal adenocarcinoma epithelial cell, Left panel), A431 (human epidermoid carcinoma epithelial cell, Right panel) cells labelling EGFR with ab264540 at 1/100 dilution (1µg) (Red) compared with a mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Negative control: SW620.
Gated on viable cells.
This image was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (ab264540).
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Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling EGFR with ab264540 at 1/100 dilution followed by a ready to use secondary. Membranous staining on human lung carcinoma tissue is observed. The section was incubated with ab264540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This image was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (ab264540).
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Immunohistochemical analysis of paraffin-embedded human oral carcinoma tissue labeling EGFR with ab264540 at 1/100 dilution followed by a ready to use secondary. Membranous staining on human oral carcinoma tissue is observed. The section was incubated with ab264540 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This image was produced using the same antibody clone but in a different formulation containing PBS, sodium azide, glycerol and BSA (ab264540).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (2)
ab264547 has been referenced in 2 publications.
- Kaneko MK et al. Elucidation of the critical epitope of an anti-EGFR monoclonal antibody EMab-134. Biochem Biophys Rep 14:54-57 (2018). PubMed: 29872734
- Itai S et al. Establishment of EMab-134, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody for Detecting Squamous Cell Carcinoma Cells of the Oral Cavity. Monoclon Antib Immunodiagn Immunother 36:272-281 (2017). PubMed: 29090976