Phospho-EGFR (Y992) Recombinant Rabbit Monoclonal Antibody [PSH23-63]
cat.: HA751904
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH23-63
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 134 kDa
Isotype: IgG
Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Tyr992 of human EGFR.
Positive control: A431 treated with 100ng/mL EGF for 30min cell lysate.
Subcellular location: Cell membrane, Nucleus membrane, Nucleus, Endoplasmic reticulum membrane, Golgi apparatus membrane, Endosome.
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:2,000
1:1,000
1:10,000
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
HA751904_1.jpg Fig1: Western blot analysis of Phospho-EGFR (Y992) on different lysates with Rabbit anti-Phospho-EGFR (Y992) antibody (HA751904) at 1/2,000 dilution.

Lane 1: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
Lane 2: A431 treated with 100ng/mL EGF for 30min cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 12 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751904, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 134 kDa
Observed band size: 175 kDa
HA751904_2.jpg Fig2: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cells) treated with EGF(100ng/ml 30min)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751904, 1/1,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
HA751904_3.jpg Fig3: Application: Flow Cytometry (Intra)

Species: Human
Sample: A431 (Human epidermoid carcinoma skin squamous cells) treated with EGF(100ng/ml 30min)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751904 (1/10,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.