Anti-EGFR antibody
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概述
- 产品描述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 cascad.
- 产品名称Anti-EGFR antibody
- 分子量175 kDa
- 种属反应性Human,Mouse,Rat
- 验证应用WB,ICC,IHC-P,FC
- 抗体类型兔多抗
- 免疫原This antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to a region of N-terminal residues of mouse EGFR.
- 偶联Non-conjugated
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性能
- 形态Liquid
- 浓度1 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, 40% Glycerol. Preservative: 0.05% Sodium Azide.
- 亚型IgG
- 纯化方式Peptide affinity purified.
- 亚细胞定位Plasma membrane, Nucleus, Endosome, Endoplasmic reticulum.
- 数据链接SwissProt: P00533 Human
SwissProt: Q01279 Mouse
Entrez Gene: 24329 Rat
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wa2 antibody
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应用
WB:1:500-1:1,000
ICC:1:50-1:200
IHC-P:1:50-1:200
FC:1:50-1:100
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Fig1: Western blot analysis of EGFR on A431 cell lysate. 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 at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Fig2: ICC staining EGFR in A172 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 antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
Fig3: Immunohistochemical analysis of paraffin-embedded rat stomach tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 antibody at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Fig4: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-EGFR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 antibody t 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained 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 EGFR antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for at least 30 minutes.
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