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Anti -ErbB2, phosphorylated, Tyr1248 (c-erbB2, HER2, neu Oncoprotein)
Anti -ErbB2, phosphorylated, Tyr1248 (c-erbB2, HER2, neu Oncoprotein)
Pricing
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| Clone |
Host |
Grade |
Applications |
| Polyclonal |
Rabbit |
Affinity Purified |
B |
|
| Catalog # | E3451-25 | | Applications | Suitable for use in Western Blot. Other applications not tested. | | Recommended Dilution | Western Blot: 1:2000 -1:8000 dilutions detects activated Erb B2/HER-2 in RIPA lysates from heregulin stimulated MCF-7 breast carcinoma cells. Lysates from non-stimulated and heregulin-stimulated MCF-7 cells were resolved by electrophoresis, transferred to nitrocellulose and probed with E3451-25 (1:2000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. | | Optimal dilution determined by the researcher. | | Immunoblot Protocol 1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell lysate sample (cell lysis buffer | 50mM Tris-HCl, pH7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1ug/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. 2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk (PBS-MLK) for 30 minutes at RT with constant agitation. 3. Incubate the nitrocellulose with a 1:2000-1:8000 dilution of E3451-25 diluted in freshly prepared PBS-MLK overnight with agitation at 4°C. 4. Wash the nitrocellulose twice with water. 5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-rabbit HRP conjugated IgG, 1:5000 dilution was used) in PBS-MLK for 1.5 hours at room temperature with agitation. 6. Wash the nitrocellulose with water twice. 7. Wash the nitrocellulose in PBS-0.05% Tween 20 for 3-15 minutes. 8. Rinse the nitrocellulose in water for 30-60min. 9. Use detection method of choice (enhanced chemiluminescence was used). | | Storage and Stability | May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. | | CAS Number | n/a | | Clone Type | Polyclonal | | Isotype | IgG | | Host | Rabbit | | Source | Human | | Concentration | Not determined | | Form | Supplied as a liquid in 0.2M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide. | | Purity | Purified by immunoaffinity chromatography. | | Immunogen | Synthetic phosphopeptide, residues 1243-1255, corresponding to a major auto-phosphorylation (Tyr1248) site of human Erb B2/HER-2 (C-AENPEpYLGLDVPV). | | Specificity | Recognizes the activated form of human Erb B2/HER-2 oncoprotein at 185 kD, phosphorylated at Tyr1248. Species Crossreactivity: human. Predicted to crossreact with rat based on conservation of immunogenic sequence. | | | Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
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