Phospho-EGFR (Y1110) Recombinant Rabbit Monoclonal Antibody [PSH27-58]
cat.: HA724705
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH27-58 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| 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 Tyr1110 of Human EGFR. |
| Positive control: | A431 treated with 100ng/mL EGF for 30 minutes cell lysate. |
| Subcellular location: | Cell membrane, Nucleus membrane, Nucleus, Endoplasmic reticulum membrane, Golgi apparatus membrane. |
| Recommended Dilutions:
WB IF-Cell IP |
1:5,000 1:500 1-2μg/sample |
| 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
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Fig1:
Western blot analysis of Phospho-EGFR (Y1110) on different lysates with Rabbit anti-Phospho-EGFR (Y1110) antibody (HA724705) at 1/5,000 dilution.
Lane 1: A431 (Human epidermoid carcinoma skin squamous cell) cell lysate Lane 2: A431 treated with 100ng/mL EGF for 30 minutes cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 42 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724705, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C 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 |
|
Fig2:
Application: Immunocytochemistry (IF-cell)
Species: Human Sample: A431 (Human epidermoid carcinoma skin squamous cell) treated with or without 100ng/mL EGF for 30 minutes 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: HA724705, 1/500, 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). |
|
Fig3:
Immunoprecipitation (IP)
Phospho-EGFR (Y1110) was immunoprecipitated in 0.2 mg A431 (Human epidermoid carcinoma skin squamous cell) treated with 100ng/mL EGF for 30 minutes cell lysate with HA724705 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724705 at 1/5,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A431 treated with 100ng/mL EGF for 30 minutes cell lysate (input) Lane 2: HA724705 IP in A431 treated with 100ng/mL EGF for 30 minutes cell lysate Lane 3: Rabbit IgG instead of HA724705 in A431 treated with 100ng/mL EGF for 30 minutes cell lysate Exposure time: 30 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA724705, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 134 kDa Observed band size: 175 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.