Mouse Recombinant Monoclonal EGFR antibody. Carrier free. Suitable for ICC/IF, WB, IP and reacts with Human samples.
IgG1
Mouse
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
ICC/IF | WB | IHC-P | Flow Cyt (Intra) | IP | |
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Human | Tested | Tested | Not recommended | Not recommended | Tested |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Mouse, Rat | Dilution info - | Notes - |
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Species Mouse, Rat, Human | Dilution info - | Notes - |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
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Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses (PubMed:2790960, PubMed:10805725, PubMed:27153536). Known ligands include EGF, TGFA/TGF-alpha, AREG, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF (PubMed:2790960, PubMed:7679104, PubMed:8144591, PubMed:9419975, PubMed:15611079, PubMed:12297049, PubMed:27153536, PubMed:20837704, PubMed:17909029). 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).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.
Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1, EGFR, ERBB, ERBB1, HER1
Mouse Recombinant Monoclonal EGFR antibody. Carrier free. Suitable for ICC/IF, WB, IP and reacts with Human samples.
Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1, EGFR, ERBB, ERBB1, HER1
IgG1
Mouse
pH: 7.2 - 7.4
Constituents: 100% PBS
Liquid
Monoclonal
Yes
13/EGFR
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab289896 is the carrier free version of Anti-EGFR antibody [13/EGFR] ab289889.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was developed using Anti-EGFR antibody [13/EGFR] ab289889, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
False colour image of Western blot: Anti-EGFR antibody [13/EGFR] (Anti-EGFR antibody [13/EGFR] ab289889 staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red.
In Western blot, Anti-EGFR antibody [13/EGFR] ab289889 was shown to bind specifically to EGFR. A band was observed at 170 kDa in wild-type HeLa cell lysates with no signal observed at this size in the EGFR knockout cell line. To generate this image, wild-type and EGFR knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1 % TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution.
Performed under reducing conditions.
Low expression: SW480(PMID: 19789347)
We observe an unknown band at around 38k.
All lanes: Western blot - Anti-EGFR antibody [13/EGFR] (Anti-EGFR antibody [13/EGFR] ab289889) at 1/1000 dilution
Lane 1: Wild type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: EGFR knockout HeLa whole cell lysate at 20 µg
Lane 3: NCI-H1975 (Human adenocarcinoma lung epithelial cell) whole cell lysate at 20 µg
Lane 4: SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
All lanes: Goat Anti-Mouse IgG H&L (IRDye® 800CW) (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 134 kDa
Observed band size: 170 kDa
This data was developed using Anti-EGFR antibody [13/EGFR] ab289889, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-431 (Human epidermoid carcinoma cell line) labeling EGFR with Anti-EGFR antibody [13/EGFR] ab289889 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) preadsorbed secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in A431 cell line.
Low expression:SW480 (PMID: 27729020). The nuclear counter stain is DAPI (blue).
Tubulin is counterstained with Anti-alpha Tubulin mouse MAb (Anti-beta Tubulin antibody [EPR16774] ab179513) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-EGFR antibody [13/EGFR] ab289889 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-beta Tubulin antibody [EPR16774] ab179513) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) secondary antibody at 1/1000 dilution.
This data was developed using Anti-EGFR antibody [13/EGFR] ab289889, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-EGFR antibody [13/EGFR] (Anti-EGFR antibody [13/EGFR] ab289889) at 1/1000 dilution
All lanes: A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 10 µg
All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 134 kDa
Observed band size: 170 kDa
Exposure time: 15s
This data was developed using Anti-EGFR antibody [13/EGFR] ab289889, the same antibody clone in a different buffer formulation.
EGFR was immunoprecipitated from A431 (human epidermoid carcinoma epithelial cell) whole cell lysate 10 μg with Anti-EGFR antibody [13/EGFR] ab289889 at 1/30 diultion. Western Blot was performed from immunoprecipitate using Anti-EGFR antibody [13/EGFR] ab289889 at 1/1000 diultion. Mouse IgG for IP (HRP) (Anti-mouse IgG for IP (HRP) ab131368) was used as a secondary antibody at 1/5000 dilution.
Lane 1 (Input): A431 (human epidermoid carcinoma epithelial cell) whole cell lysate)
Lane 2 (+): A431 whole cell lysate
Lane 3 (-): Mouse IgG1 monoclonal (Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443) instead of Anti-EGFR antibody [13/EGFR] ab289889 in A431 whole cell lysate
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-EGFR antibody [13/EGFR] (Anti-EGFR antibody [13/EGFR] ab289889)
Predicted band size: 134 kDa
Exposure time: 3min
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