Anti-EGFR antibody [7-F2-F8]

Anti-EGFR antibody [7-F2-F8]

• 概述

• 产品描述The EGF receptor family comprises several related receptor tyrosine kinases that are frequently overexpressed in a variety of carcinomas. Members of this receptor family include EGFR (HER1), Neu (ErbB-2, HER2), ErbB-3 (HER3) and ErbB-4 (HER4), which form either homodimers or heterodimers upon ligand binding. EGFR binds several ligands, including epidermal growth factor (EGF), transforming growth factor α (TGFα), Amphiregulin and heparin binding-EGF (HB-EGF). Ligand binding promotes the internalization of EGFR via Clathrin-coated pits and its subsequent degradation in response to its intrinsic tyrosine kinase. EGFR is involved in organ morphogenesis and maintenance and repair of tissues, but upregulation of EGFR is associated with tumor progression. The oncogenic effects of EGFR include initiation of DNA synthesis, enhanced cell growth, invasion and metastasis. Abrogation of EGFR results in cell cycle arrest, apoptosis or dedifferentiation of cancer cells, suggesting that EGFR may be an effective therapeutic target.
• 产品名称Anti-EGFR antibody [7-F2-F8]
• 分子量Predicted band size 134 kDa.
• 种属反应性Human
• 验证应用WB,ICC,IHC-P,FC
• 抗体类型小鼠单抗
• 免疫原Recombinant protein within human EGFR aa 900-1150.
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度2 mg/mL.
• 存放说明Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型IgG2b
• 纯化方式Protein A affinity purified.
• 亚细胞定位Golgi apparatus membrane, nucleus membrane, nucleus, cell membrane, endosome, endosome membrane, endoplasmic reticulum membrane.
• 其它名称
  • Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
  • Cell growth inhibiting protein 40 antibody
  • Cell proliferation inducing protein 61 antibody
  more

• 应用

  WB:1:500-1:2,000
  ICC:1:50-1:100
  IHC-P:1:50-1:200
  FC:1:50-1:100

•  

  

  Fig1: Western blot analysis of EGFR on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  Positive control:
  Lane 1: A431 cell lysate
  Lane 2: SKOV-3 cell lysate
  Lane 3: A549 cell lysate

   

  

  Fig2: ICC staining of EGFR in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

   

  

  Fig3: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-67, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig5: Flow cytometric analysis of EGFR was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody  (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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