Product Name:
EGFR-pY998
Product Number:
ab-pk603
Target Full Name: Epidermal growth factor receptor-tyrosine kinase
Target Alias: EGFR; Epidermal growth factor receptor; ErbB-1; ErbB, mENA; HER1; Receptor tyrosine-protein kinase ErbB-1; V-erb-b oncogene homologue; PIG61; CCDS5514.1; ENSG00000146648
Product Type Specific: EGFR protein kinase phosphosite-specific antibody
Antibody Code: PK603
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y998
Protein UniProt: P00533
Protein SigNET: EGFR
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human EGFR (ErbB1) sequence peptide Cat. No.: PE-04ACR99
Antibody Immunogen Sequence: SNF(pY)RAL(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues S995 to L1001; in the cytoplasmic region of EGFR after the kinase catalytic domain. Y998 phosphorylation inhibits phosphotransferase activity. Phosphorylation induces molecular association with CBL, CRK, CSK, SHC1, SHP-2 and STAT5A, as well as receptor internalization. This is one of the major in vivo phosphorylation sites in EGFR (≥147 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.75 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Very weak immunoreactivity of a target-sized protein by Western blotting in A431 cells (homogenized in SDS-PAGE sample buffer). Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Dog | Mouse | Platypus.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 160-180 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431 cells, except for a ~50 kDa cross-reactive protein.
Related Product 1: EGFR (995-1001) pY998 antibody blocking peptide (Cat. No.: PE-04ACR99)
Related Product 2: EGFRSubtide kinase substrate peptide (Cat. No.: PE-01BHE95)
Related Product 3: EGFR T678 phosphorylation antibody (Cat. No.: AB-PK120)
Related Product 4: EGFR T693 phosphorylation antibody (Cat. No.: AB-PK121)
Related Product 5: EGFR Y869 phosphorylation antibody (Cat. No.: AB-PK602)
Related Product 6: EGFR Y1069 phosphorylation antibody (Cat. No.: AB-PK599)
Related Product 7: EGFR Y1110 phosphorylation antibody (Cat. No.: AB-PK600)
Related Product 8: EGFR Y1172 phosphorylation antibody (Cat. No.: AB-PK601)
Related Product 9: EGFR expression antibody (Cat. No.: AB-NK052-6)
Related Product 10: EGFR expression antibody (Cat. No.: AB-NK052-4)
Scientific Background: EGFR (ERBB1, Erbb1, HER1) is a protein-tyrosine kinase of the TK group and EGFR family. It features three main domains: an extracellular ligand-binding domain (ECD), a transmembrane domain, and an intracellular tyrosine kinase domain. EGFR regulates cell proliferation, division, motility, survival, and has other roles in tissue development. Although the encoded protein is 134 kDa based on its amono acid sequence, EGFR undergoes extensive post-translational modifications, including phosphorylation and glycosylation, which affects its measured mass on SDS-PAGE gels (typically 170-180 kDa. It is activated by binding epidermal growth factor (EGF) and transforming growth factor-alpha (TGFa), which induce dimerization and autophosphorylation. Phosphorylation of Tyr-270 stimulates interaction with EGFR. Phosphorylation of Tyr-869 increases phosphotransferase activity and induces interaction with COX2. Phosphorylation of Tyr-1016 increases phosphotransferase activity and induces interaction with GrbB2, PLCg1, PTPN11 (SHP2), RasGAP, and Vav2. Phosphorylation of Tyr-1069 increases phosphotransferase activity and induces interaction with Cbl. Phosphorylation of Y1092 induces interaction with PLCg1, PTPN6 (SHP1) and Ras-GAP. Phosphorylation of Y1110 induces interaction with Ras-GAP. Phosphorylation of Tyr-1172 increases phosphotransferase activity and induces interaction with Dok1, EGFR, RasGAP, PTPN11 (SHP2), and Vav2. Phosphorylation of Tyr-1197 increases phosphotransferase activity and induces interaction with Cbl, EGR, PLCg1, RasGAP, SH3KBP1, Shc1, and PTPN6 (SHP1). EGFR is inhibited by phosphorylation at Thr-678, Ser-695 and Ser-1026. Phosphorylation of Thr-693, Ser-695, Ser-1070, Ser-1071, and Ser-1190 contributes to receptor internalization. EGFR is found on the plasma membrane, endosomes, and sometimes in the nucleus. It is a known oncoprotein (OP). Cancer-related mutations in human tumours point to a gain of function of the protein kinase. Constitutive activation of EGFR kinase activity has been seen with the mutations of V689M, E1005R+D1006K. Overexpression of EGFR, particularly in non-small cell lung cancer (NSCLC), lead to uncontrolled cell division. The active form of the protein kinase normally promotes tumour cell proliferation. This description may include information annotated by UniProt and/or Google AI.