EGFR (C-term) (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 13G8]

CAT#: AM00047PU-N

EGFR (C-term) (incl. pos. control) mouse monoclonal antibody, clone 13G8, Purified

Product Images

Detection of endogenous EGFR: Whole cell lysates of serum starved tumor cells (20.000 cells per lane) were applied to SDS-PAGE and transferred to a PVDF membrane. The immunoblot was probed with mab EGFR-13G8 (0.5 ug/ ml) for 1h at RT and developed by ECL (exp. time: 30 sec). lane 1: A431; lane 2: A549; lane 3: SKOV3; lane 4: OVCAR5; lane 5: HaCaT; lane 6: PC3; lane 7: HeLa; lane 8: HepG2

Specifications

Product Data

• Clone Name: 13G8
• Applications: ELISA, IF, IP, WB
• Recommended Dilution: ELISA (0.05 µg/ml).
  Immunoblotting: 1 μg/ml for HRPO/ECL detection.
  Recommended blocking buffer: Casein/Tween 20 based blocking and blot incubation buffer AS00002BU-N or AS00002BU-L.
  Included Positive Control: Cell lysate from untreated HepG2 cells (See Protocols for more details).
  Immunoprecipitation (1-10 µg/ml per 10e6 pervanadate-treated A431 cells).
  Immunocytochemistry (1-10 µg/ml).
• Reactivities: Human, Mouse
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Peptide conjugated to KLH.
  Epitope: C-terminus (aa 1165-1186) independent of phosphorylation status.
• Specificity: This antibody specifically recognizes the C-terminus of EGF receptor (aa 1165 - 1186).
  Recognition is independent of the phosphorylation status at tyrosine 1173.
• Formulation: PBS, 0.09% Sodium Azide/PEG and Sucrose
  State: Purified
  State: Lyophilized purified IgG fraction.
• Reconstitution Method: Restore with 1 ml H2O (15 min, RT).
• Purification: Subsequent Ultrafiltration and Size Exclusion Chromatography
• Conjugation: Unconjugated
• Storage: Store lyophilized (preferably in a desiccator) at -20°C and reconstituted (aliquote and
  freeze in liquid nitrogen) at -80°C.
  Avoid repeated freezing and thawing.
  Thaw aliquots at 37°C.
  Thawed aliquots may be stored at 4°C up to 3 months.
• Stability: Shelf life: one year from despatch.
• Gene Name: epidermal growth factor receptor
• Database Link:
  Entrez Gene 1956 Human
  UniProt ID P00533
• Background: EGFR/erbB receptors are activated upon binding of EGF and EGF-related growth factors such as TGF alpha, beta-cellulin, Hb-EGF, HRG, or NRG. Binding of these ligands leads to receptor homo- and heterodimerization followed by autophosphorylation and activation of downstream signal transduction pathways (MAPK, PI3K/PKB, and STAT). In addition, EGFR becomes fully activated after phosphorylation of Y845 by src family kinases.
  Phosphorylation of Y1045 leads to association with cbl and subsequent receptor degradation. Phosphorylation of S1047 by CamKinase II leads to attenuation of kinase activity; phosphorylation of T654 (by PKC) and T669 (by MAPK, p38) interferes with receptor endocytosis/recycling.
• Synonyms: Epidermal growth factor receptor, EGF Receptor, erbB-1, c-ErbB-1
• Note: Mol. weight: 180 kDa.
  
  Protocol: Positive Control Provided: Cell lysate from untreated HepG2 cells.
  
  Description: Cell lysate from untreated HepG2 cells.
  
  Format: Lyophilized cell lysate from serum starved HepG2 cells.
  
  Reconstitution: Restore by addition of 200 μl H20. After complete solubilization add 200 μl
  2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min.
  
  Storage: Aliquote and store frozen.
  Avoid repeated freeze/thaw cycles.
  
  Application: The positive control cell lysate is recommended for immunoblot applications.
  20 μl of positive control cell lysate correspond to ca. 80.000 cells. Use 20 μl / lane (mini
  gel) for HRPO/ECL detection of the target proteins.
  Please note: The lyophilized cell lysates contain SDS and are not recommended for
  applications with native proteins such as immunoprecipitation.