Recombinant Alexa Fluor® 647 Anti-EGFR antibody [EP38Y] (ab192982)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Alexa Fluor® 647 Rabbit monoclonal [EP38Y] to EGFR
- Suitable for: ICC/IF, Flow Cyt (Intra)
- Reacts with: Human
- Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
Related conjugates and formulations
Overview
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Product name
Alexa Fluor® 647 Anti-EGFR antibody [EP38Y]
See all EGFR primary antibodies -
Description
Alexa Fluor® 647 Rabbit monoclonal [EP38Y] to EGFR -
Host species
Rabbit -
Conjugation
Alexa Fluor® 647. Ex: 652nm, Em: 668nm -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC/IF: HeLa and A431 cells, Flow Cyt (intra): HeLa
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. Store In the Dark. -
Dissociation constant (KD)
KD = 1.90 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol (glycerin, glycerine), 1% BSA, PBS -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Monoclonal -
Clone number
EP38Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-EGFR antibody [EP38Y] - Low endotoxin, Azide free (ab174481)
- Alexa Fluor® 488 Anti-EGFR antibody [EP38Y] (ab193244)
- HRP Anti-EGFR antibody [EP38Y] (ab193602)
- Alexa Fluor® 594 Anti-EGFR antibody [EP38Y] (ab207870)
- PE Anti-EGFR antibody [EP38Y] (ab208753)
- Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)
- Alexa Fluor® 555 Anti-EGFR antibody [EP38Y] (ab274892)
- Biotin Anti-EGFR antibody [EP38Y] (ab314262)
- Anti-EGFR antibody [EP38Y] (ab52894)
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192982 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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ICC/IF |
1/50 - 1/1000.
This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min) and A431 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min). |
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Flow Cyt (Intra) |
Use 2µl for 106 cells.
ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/50 - 1/1000. This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min) and A431 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min). |
Flow Cyt (Intra)
Use 2µl for 106 cells. ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody. |
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
- Entrez Gene: 13649 Mouse
- Entrez Gene: 24329 Rat
- Omim: 131550 Human
- SwissProt: P00533 Human
- SwissProt: Q01279 Mouse
- Unigene: 488293 Human
- Unigene: 605083 Human
see all -
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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ab192982 staining EGFR in A431 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab192982 at 1/1000 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in A431 cells fixed with 100% methanol (5 min).
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ab192982 staining EGFR in HeLa cells. The cells were fixed with 100% methanol (5 min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton X-100 for 1hr. The cells were then incubated with ab192982 at a working dilution of 1 in 50 (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1hr with AlexaFluor® 488 Goat anti-mouse IgG (H&L - preadsorbed) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a Confocal microscope (Leica-microsystems, TCS SP8).
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Overlay histogram showing HeLa cells stained with ab192982 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab192982, 2μl/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa Fluor® 647) (1μg/1x106 cells) for 30 min at 22ºC. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 25mW red solid state diode laser (635nm) and 675/30 bandpass filter.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab192982 has been referenced in 3 publications.
- Deng H et al. A fluorogenic probe for predicting treatment response in non-small cell lung cancer with EGFR-activating mutations. Nat Commun 13:6944 (2022). PubMed: 36376325
- Fu C et al. EWI-2 controls nucleocytoplasmic shuttling of EGFR signaling molecules and miRNA sorting in exosomes to inhibit prostate cancer cell metastasis. Mol Oncol 15:1543-1565 (2021). PubMed: 33605506
- Eng J et al. Cyclic Multiplexed-Immunofluorescence (cmIF), a Highly Multiplexed Method for Single-Cell Analysis. Methods Mol Biol 2055:521-562 (2020). PubMed: 31502168