Phospho-EGFR (Y1092) Recombinant Rabbit Monoclonal Antibody [SJ0194]
cat.: ET1606-44
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SJ0194
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: 170 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Tyr1092 of Human EGFR aa 1,071-1,120 / 1,210.
Positive control: A431 cell lysate treated with EGF, BT-20, human tonsil tissue, mouse skin tissue.
Subcellular location: Cell membrane, Nucleus membrane, Nucleus, Endoplasmic reticulum membrane, Golgi apparatus membrane, Endosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
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
ET1606-44_1.jpg Fig1: Western blot analysis of Phospho-EGFR (Y1092) 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 (ET1606-44, 1/1,000) was used 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.

Positive control:
Lane 1: A431 cell lysate treated with EGF
Lane 2: Untreated A431 cell lysate
ET1606-44_2.jpg Fig2: Western blot analysis of Phospho-EGFR(Y1092) on A431 cell lysates.

Lane 1: A431 cells, whole cell lysate, 10ug/lane
Lane 2/3: A431 cells treated with 100 ng/ml EGF for 30 minutes, whole cell lysates, 10ug/lane
Lane 4: A431 cells treated with 100 ng/ml EGF for 30 minutes, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane

All lanes :
Anti-Phospho-EGFR(Y1092) antibody (ET1606-44
) at 1:500 dilution. Anti-EGFR antibody (ET1603-37) at 1:500 dilution. Anti-Hsp90 beta antibody (ET1605-56) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.

Predicted band size: 134 kDa
Observed band size: 170 kDa

Blocking and diluting buffer: 5% BSA.

Exposure time: 15 seconds
ET1606-44_3.jpg Fig3: ICC staining of Phospho-EGFR (Y1092) in BT-20 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 (ET1606-44, 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).
ET1606-44_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-EGFR (Y1092) 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 (ET1606-44, 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.
ET1606-44_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-Phospho-EGFR (Y1092) 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 (ET1606-44, 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.
ET1606-44_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Phospho-EGFR (Y1092) antibody (ET1606-44) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-44) at 1/200 dilution for 1 hour 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”.