Phospho-EGFR (Y1173) Recombinant Rabbit Monoclonal Antibody [SC57-04]
cat.: ET1610-4
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | SC57-04 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 134 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Tyr1173 of Human EGFR. |
| Positive control: | A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate, A431, B16F1, human placenta tissue, mouse skin tissue. |
| Subcellular location: | Cell membrane, Nucleus membrane, Nucleus, Endoplasmic reticulum membrane, Golgi apparatus membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P IP |
1:1,000-1:2,000 1:50-1:200 1:50-1:200 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P00533 Human | Q01279 Mouse |
| Alternative names: | Avian erythroblastic leukemia viral (v erb b) oncogene homolog Cell growth inhibiting protein 40 Cell proliferation inducing protein 61 EGF R EGFR EGFR_HUMAN Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) Epidermal growth factor receptor (erythroblastic leukemia viral (v erb b) oncogene homolog avian) Epidermal growth factor receptor erb-b2 receptor tyrosine kinase 1 ERBB ERBB1 Errp HER1 mENA NISBD2 Oncogen ERBB PIG61 Proto-oncogene c-ErbB-1 Receptor tyrosine protein kinase ErbB 1 Receptor tyrosine-protein kinase ErbB-1 SA7 Species antigen 7 Urogastrone v-erb-b Avian erythroblastic leukemia viral oncogen homolog wa2 Wa5 |
Images
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Fig1:
Western blot analysis of Phospho-EGFR (Y1173) on different lysates with Rabbit anti-Phospho-EGFR (Y1173) antibody (ET1610-4) at 1/2,000 dilution. Lane 1: A431 whole cell lysate Lane 2: A431 treated with 100ng/mL EGF for 30 minutes whole cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 135 kDa Observed band size: 170 kDa Exposure time: 2 minute; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-4) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/200,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of A431 cells treated with or without 100ng/mL EGF for 5 minutes labeling Phospho-EGFR (Y1173) with Rabbit anti-Phospho-EGFR (Y1173) antibody (ET1610-4) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS permeabilization for 5 minutes, and then blocked with 2% negative goat serum for 60 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-EGFR (Y1173) antibody (ET1610-4) at 1/50 dilution in 1% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. |
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Fig3: ICC staining of Phospho-EGFR (Y1173) in untreated B16F1 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 (ET1610-4, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Phospho-EGFR (Y1173) 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 primary antibody (ET1610-4, 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. |
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Fig5: Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-Phospho-EGFR (Y1173) 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 primary antibody (ET1610-4, 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.