EGFR Monoclonal Antibody (H11)/EGFR单克隆抗体（H11）

货号：MA5-13070

规格：500 µL

价格：5548

产品类型：免疫组化一抗

品牌：Thermo Fisher

抗原：HC2 20 d2 cells

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG1, kappa

克隆号：H11

荧光染料：其它

类型:	单抗	同型对照：
浓度：	0.2 mg/mL	用法：	0.5-1 µg/test(Flow)；2 µg/mL(ICC)；2 µg/mL(IF)；2-4 µg/mL(IHC (P))；2 µg/mL(IP)；0.5-1.0 µg/mL(WB)

产品详细信息MA5-13070 targets Epidermal Growth Factor Receptor in FACS, ICC/IF, IHC (P), IP, and WB applications and shows reactivity with Human and Mouse samples. This antibody is not recommended for mouse lymph node tissue or human breast carcinoma in IHC applications.The MA5-13070 immunogen is hC2 20 d2 cells.靶标信息EGFR (Epidermal growth factor receptor, HER1, ErbB1) is encoded by the EGFR gene located on chromosome 7 in humans. EGFR belongs to the HER/ERbB family of proteins that includes three other receptor tyrosine kinases, ERbB2, ERbB3, ERbB4. EGFR is a transmembrane receptor and binding of its cognate ligands such as EGF (Epidermal Growth Factor) and TGF alpha (Transforming Growth Factor alpha) to the extracellular domain leads to EGFR dimerization followed by autophosphorylation of the tyrosine residues in the cytoplasmic domain. Phosphorylation of EGFR at certain residues is also mediated by Src-non-receptor kinase. EGFR activation signals multiple downstream signaling cascades such as the Ras - ERK, PI3-K - Akt, Jak - STAT and PKC pathways that help in growth and proliferation of cells. Phosphorylation of EGFR at Y1086 specifically allows binding of the adaptor protein GRB2, leading to activation of the MAPK pathway. Upon receptor activation and signaling, EGFR is endocytosed and targeted for degradation or recycling. Mutations in the EGFR gene are associated with lung cancer and multiple alternatively spliced transcript variants encode different protein isoforms of EGFR have been found. Increased production or activation of EGFR has been associated with poor prognosis in a variety of tumors. Moreover, EGFR overexpression is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.仅用于科研。不用于诊断过程。未经明确授权不得转售。

数据

EGFR Antibody (MA5-13070) in IFImmunofluorescence analysis of EGFR was performed using 90% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EGFR Mouse monoclonal antibody (Product # MA5-13070) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification. EGFR Antibody (MA5-13070) in IFImmunofluorescent analysis of Epidermal Growth Factor Receptor (green) showing staining in the membrane of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with an Epidermal Growth Factor Receptor monoclonal antibody (Product # MA5-13070) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x. EGFR Antibody (MA5-13070) in IHC (P)Immunohistochemistry analysis of EGFR showing staining in the membrane and cytoplasm of paraffin-treated human lung carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Epidermal Growth Factor Receptor monoclonal antibody (Product # MA5-13070) diluted by 3% BSA-PBS at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

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参考文献：

1 Nature communicationsSortilin limits EGFR signaling by promoting its internalization in lung cancer.2 OncotargetMUC1 stimulates EGFR expression and function in endometrial cancer.3 OdontologyCoexpression of SGLT1 and EGFR is associated with tumor differentiation in oral squamous cell carcinoma.4 The Journal of biological chemistryPhysical interaction between epidermal growth factor receptor and DNA-dependent protein kinase in mammalian cells.5 OncogeneActivity and cellular localization of an oncogenic glioblastoma multiforme-associated EGF receptor mutant possessing a duplicated kinase domain.

技术参数

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