Anti-EGFR Affibody® Molecule (ab95116)
Overview
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Product name
Anti-EGFR Affibody® Molecule
See all EGFR affibody® molecule -
Specificity
The Anti-EGFR Affibody® molecule binds to human EGFR. Cross reactivity with other species has not been tested. -
Tested applications
Suitable for: ICC/IF, IHC-Fr, Flow Cytmore details -
Species reactivity
Reacts with: Human -
General notes
The theoretical pI of ab95116 is 4.4 and the molecular weight is 13.9kDa.A unique cysteine has been introduced at the C-terminus on a genetic level. This residue can advantageously be used for directed conjugations.
This product contains a unique C-terminal cysteine and spontaneously generates tail-to-tail dimers via a disulfide bridge between the C-terminal cysteines. Depending on the storage conditions and handling, the amount of dimers may increase over time. Prior to coupling via the C-terminal cysteine or chemical modification directed to the unique cysteine residue, this Affibody? molecule needs to be reduced to expose a reactive cysteine residue. Recommended reducing condition is 20 mM DTT at a pH above 7.5 for incubation at room temperature for approximately 1 hour. Immediately prior to coupling or modification of the cysteine, excess DTT should be removed by passage through a desalting column. As the Affibody® molecule will dimerize quickly, dialysis is not recommended.
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. -
Storage buffer
pH: 7.40
Constituents: 0.079% Ammonium bicarbonate, PBS -
Concentration information loading... -
Purification notes
Purity: >98% by SDS-PAGE and RP-HPLC. -
Affibody® molecule notes
A unique cysteine has been introduced at the C-terminus on a genetic level. This residue can advantageously be used for directed conjugations.
This product contains a unique C-terminal cysteine and spontaneously generates tail-to-tail dimers via a disulfide bridge between the C-terminal cysteines. Depending on the storage conditions and handling, the amount of dimers may increase over time. Prior to coupling via the C-terminal cysteine or chemical modification directed to the unique cysteine residue, this Affibody? molecule needs to be reduced to expose a reactive cysteine residue. Recommended reducing condition is 20 mM DTT at a pH above 7.5 for incubation at room temperature for approximately 1 hour. Immediately prior to coupling or modification of the cysteine, excess DTT should be removed by passage through a desalting column. As the Affibody® molecule will dimerize quickly, dialysis is not recommended. -
Research areas
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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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Alternative names
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog
- Cell growth inhibiting protein 40
- Cell proliferation inducing protein 61
see all -
Database links
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
Images
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SDS-PAGE - Anti-EGFR Affibody® Molecule (ab95116)SDS-PAGE analysis of purified anti-EGFR Affibody molecule at reduced condition (lane 1) and at nonreduced condition (lane 2) on NuPage 4-12 % Bis-Tris gel. Coomassie blue was used for staining
