Phospho-EGFR (Y1068) Recombinant Rabbit Monoclonal Antibody [SD2055]
cat.: ET1612-30
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IP, FC
Clonality: Monoclonal
Clone number: SD2055
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: 134 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Tyr1068 of human EGFR.
Positive control: HeLa cell lysate, HeLa treated with 100ng/mL EGF for 30 minutes cell lysate, A431, Hela, MCF-7.
Subcellular location: Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus membrane, Nucleus membrane, Endosome, Nucleus, Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  FC
  IP
1:500-1:5,000
1:50-1:200
1:50-1:200
1:50-1:100
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
ET1612-30_1.jpg Fig1: Western blot analysis of Phospho-EGFR (Y1068) on different lysates with Rabbit anti-Phospho-EGFR (Y1068) antibody (ET1612-30) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 100ng/mL EGF for 30 minutes cell lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 24 seconds;

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 (ET1612-30) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1612-30_2.jpg Fig2: ICC staining of Phospho-EGFR (Y1068) in A431 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 (ET1612-30, 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).
ET1612-30_3.jpg Fig3: ICC staining of Phospho-EGFR (Y1068) in Hela 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 (ET1612-30, 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).
ET1612-30_4.jpg Fig4: ICC staining of Phospho-EGFR (Y1068) in MCF-7 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 (ET1612-30, 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).
ET1612-30_5.jpg Fig5: Western blot analysis of Phospho-EGFR (Y1068) on different lysates with Rabbit anti-Phospho-EGFR (Y1068) antibody (ET1612-30) at 1/5,000 dilution.

Lane 1: A431 cell lysate
Lane 2: A431 treated with 100ng/mL EGF for 30 minutes cell lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 28 seconds;

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 (ET1612-30) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1612-30_6.jpg Fig6: Flow cytometric analysis of Phospho-EGFR (Y1068) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-30, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.