Phospho-EGFR (Y1148) Rabbit Polyclonal Antibody
cat.: HA500580
| Product Type: | Rabbit polyclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell |
| Clonality: | Polyclonal |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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 Tyr1148 of human EGFR. |
| Positive control: | A431 starved overnight then treated with 20ng/mL EGF for 5 minutes cell lysate, A431 cells starved overnight then treated with 20ng/mL EGF for 5 minutes. |
| Subcellular location: | Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus membrane, Nucleus membrane, Endosome, Endosome membrane, Nucleus; Secreted. |
| Recommended Dilutions:
WB IF-Cell |
1:5,000 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
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Fig1:
Western blot analysis of Phospho-EGFR (Y1148) on different lysates with Rabbit anti-Phospho-EGFR (Y1148) antibody (HA500580) at 1/5,000 dilution. Lane 1: A431 cell lysate Lane 2: A431 starved overnight then treated with 20ng/mL EGF for 5 minutes cell lysate Lane 3: A431 starved overnight then treated with 20ng/mL EGF for 5 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 134 kDa Observed band size: 170 kDa Exposure time: 1 minute 23 seconds; ECL: K1801; 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 (HA500580) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of A431 cells untreated / starved overnight then treated with 20ng/mL EGF for 5 minutes labeling Phospho-EGFR (Y1148) with Rabbit anti-Phospho-EGFR (Y1148) antibody (HA500580) at 1/10,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-EGFR (Y1148) antibody (HA500580) at 1/10,000 dilution in 1% BSA in PBST 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. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.