Rabbit EGFR Polyclonal Antibody | anti-EGFR antibody

Phospho-EGFR (Tyr998) Antibody

Cat. Num. AAA31399

• Gene Names: EGFR; ERBB; HER1; mENA; ERBB1; PIG61
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Sheep (100%), Rabbit (100%)
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: EGFR; Polyclonal Antibody; Phospho-EGFR (Tyr998) Antibody; Avian erythroblastic leukemia viral (v erb b) oncogene homolog; Cell growth inhibiting protein 40; Cell proliferation inducing protein 61; EGF R; EGFR_HUMAN; Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog); Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian); Epidermal growth factor receptor; erb-b2 receptor tyrosine kinase 1; ERBB; ERBB1; Errp; HER1; mENA; NISBD2; Oncogen ERBB; PIG61; Proto-oncogene c-ErbB-1; Receptor tyrosine protein kinase ErbB 1; Receptor tyrosine-protein kinase ErbB-1; SA7; Species antigen 7; Urogastrone; v-erb-b Avian erythroblastic leukemia viral oncogen homolog; wa2; Wa5; anti-EGFR antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Bovine (100%), Sheep (100%), Rabbit (100%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-EGFR (Tyr998) Antibody detects endogenous levels of EGFR only when phosphorylated at Tyr998.; Tissue Specificity: Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-EGFR antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IHC: 1:50-1:200; IF/ICC: 1:100-1:500; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human EGFR around the phosphorylation site of Tyr998.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: Phosphorylated on Tyr residues in response to EGF. Phosphorylation 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 (By similarity).Palmitoylated on Cys residues by ZDHHC20. Palmitoylation inhibits internalization after ligand binding, and increases the persistence of tyrosine-phosphorylated EGFR at the cell membrane. Palmitoylation increases the amplitude and duration of EGFR signaling.Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197.
• Subunit Structure: Binding of the ligand triggers homo- and/or heterodimerization of the receptor triggering its autophosphorylation. Heterodimer with ERBB2. Forms a complex with CCDC88A/GIV (via SH2-like regions) and GNAI3 which leads to enhanced EGFR signaling and triggering of cell migration; binding to CCDC88A requires autophosphorylation of the EGFR C-terminal region, and ligand stimulation is required for recruitment of GNAI3 to the complex. Interacts with ERRFI1; inhibits dimerization of the kinase domain and autophosphorylation. Part of a complex with ERBB2 and either PIK3C2A or PIK3C2B. Interacts with GRB2; an adapter protein coupling the receptor to downstream signaling pathways. Interacts with GAB2; involved in signaling downstream of EGFR. Interacts with STAT3; mediates EGFR downstream signaling in cell proliferation. Interacts with RIPK1; involved in NF-kappa-B activation. Interacts (autophosphorylated) with CBL, CBLB and CBLC; involved in EGFR ubiquitination and regulation. Interacts with SOCS5; regulates EGFR degradation through ELOC- and ELOB-mediated ubiquitination and proteasomal degradation. Interacts with PRMT5; methylates EGFR and enhances interaction with PTPN6. Interacts (phosphorylated) with PTPN6; inhibits EGFR-dependent activation of MAPK/ERK. Interacts with COPG1; essential for regulation of EGF-dependent nuclear transport of EGFR by retrograde trafficking from the Golgi to the ER. Interacts with TNK2; this interaction is dependent on EGF stimulation and kinase activity of EGFR. Interacts with PCNA; positively regulates PCNA. Interacts with PELP1. Interacts with MUC1. Interacts with AP2M1. Interacts with FER. May interact with EPS8; mediates EPS8 phosphorylation. Interacts (via SH2 domains) with GRB2, NCK1 and NCK2. Interacts with ATX2. Interacts with GAREM1. Interacts (ubiquitinated) with ANKRD13A/B/D; the interaction is direct and may regulate EGFR internalization after EGF stimulation. Interacts with GPER1; the interaction occurs in an estrogen-dependent manner. Interacts (via C-terminal cytoplasmic kinase domain) with ZPR1 (via zinc fingers). Interacts with RNF115 and RNF126. Interacts with GPRC5A (via its transmembrane domain). Interacts with FAM83B; positively regulates EGFR inducing its autophosphorylation in absence of stimulation by EGF. Interacts with LAPTM4B; positively correlates with EGFR activation. Interacts with STX19. Interacts with CD44.
• Similarity: Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
• Subcellular Location: Cell membrane>Single-pass type I membrane protein. Endoplasmic reticulum membrane>Single-pass type I membrane protein. Golgi apparatus membrane>Single-pass type I membrane protein. Nucleus membrane>Single-pass type I membrane protein. Endosome. Endosome membrane. Nucleus.; Note: In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER (PubMed:20674546). Endocytosed upon activation by ligand (PubMed:2790960, PubMed:17182860, PubMed:27153536). Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF) (PubMed:20551055). Secreted.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Application Data

(At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

IHC (Immunohistochemistry)

(At 1/200 staining Human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

IHC (Immunohistochemistry)

(At 1/200 staining Mouse kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

IHC (Immunohistochemistry)

(At 1/200 staining Rat kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

WB (Western Blot)

(Western blot analysis of EGFR (Phospho-Tyr998) using LPS treated HeLa whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

WB (Western Blot)

(Western blot analysis of extracts from various samples, using Phospho-EGFR (Tyr998) Antibody.Lane 1: Hela cells(serum starvation treatment), blocked with antigen-specific peptides,Lane 2: Hela cells(serum starvation treatment),Lane 3: HepG2 cells(heat shock treatment).)

Related Product Information for anti-EGFR antibody

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, AREG, 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. 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. 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.

NCBI and Uniprot Product Information

• NCBI GI #: 29725609
• NCBI GeneID: 1956
• NCBI Accession #: NP_005219.2
• NCBI GenBank Nucleotide #: NM_005228.3
• UniProt Accession #: P00533; O00688; O00732; P06268; Q14225; Q68GS5; Q92795; Q9BZS2; Q9GZX1; Q9H2C9; Q9H3C9; Q9UMD7
• Molecular Weight: 134,277 Da
• NCBI Official Full Name: epidermal growth factor receptor isoform a
• NCBI Official Synonym Full Names: epidermal growth factor receptor
• NCBI Official Symbol: EGFR
• NCBI Official Synonym Symbols: ERBB; HER1; mENA; ERBB1; PIG61
• NCBI Protein Information: epidermal growth factor receptor; proto-oncogene c-ErbB-1; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; receptor tyrosine-protein kinase erbB-1; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog
• UniProt Protein Name: Epidermal growth factor receptor
• UniProt Gene Name: EGFR
• UniProt Synonym Gene Names: ERBB; ERBB1; HER1
• UniProt Entry Name: EGFR_HUMAN