Phospho-EGFR (Y1173) Recombinant Rabbit Monoclonal Antibody [SC57-04]
cat.: ET1610-4
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, IP
Clonality: Monoclonal
Clone number: SC57-04
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
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: 135 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, untreated A431, untreated A549, untreated 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
1:50-1:200
1:50-1:200
Use at an assay dependent concentration.
Uniprot #: SwissProt: P00533 Human
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
ET1610-4_1.jpg Fig1: Western blot analysis of Phospho-EGFR (Y1173) on different lysates with Rabbit anti-Phospho-EGFR (Y1173) antibody (ET1610-4) at 1/500 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: 150 kDa

Exposure time: 1 minute;

6% 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 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1610-4_2.jpg Fig2: ICC staining of Phospho-EGFR (Y1173) in untreated 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 (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).
ET1610-4_3.jpg 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).
ET1610-4_4.jpg 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.
ET1610-4_5.jpg 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.
ET1610-4_6.jpg Fig6: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.