| Catalog # | G8599-09 |
| Applications | Suitable for use in ELISA and Immunoblot. Other applications not tested. |
| Recommended Dilution | Immunoblot Analysis: 1-2ug/ml. Detects Gia2 from a rat brain microsomal preparation. Rat brain microsomal preparation was resolved by electrophoresis, transferred to nitrocellulose and probed with G8599-09 (1.5ug/ml). Proteins were visualized using a goat anti-mouse IgG labeled with HRP and a chemiluminescence detection system. |
| Optimal dilutions to be determined by the researcher. |
| Immunoblot Protocol | 1. Perform SDS-PAGE on a microsomal membrane preparation. Transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. |
| 2. Block the blotted nitrocellulose in freshly prepared PBS, 3% nonfat dry milk (PBS-MLK) for 60 minutes at RT with constant agitation. |
| 3. Incubate the nitrocellulose with 1-2ug/ml of G8599-09, diluted in freshly prepared PBS-MLK, overnight at 4°C with constant agitation. |
| 4. Wash the nitrocellulose twice with water. |
| 5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse IgG labeled with HRP was used) in PBS-MLK for 1.5 hours at RT with constant agitation. |
| 6. Wash the nitrocellulose with water twice. |
| 7. Wash the nitrocellulose in PBS, 0.05% Tween-20 for 10 minutes. |
| 8. Rinse the nitrocellulose in 4-5 changes of water for 2 hours. |
| 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, store 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 | Monoclonal |
| Isotype | IgG1 |
| Clone No | 4i41 |
| Host | Mouse |
| Source | Rat |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in 0.13M Tris, pH 7.4, 0.32M acetic acid, 30% glycerol. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | Full length, rat Gia2 puri ed by ion exchange chromatography. |
| Specificity | Recognizes rat Gia2, Mr 41kD. Broad species crossreactivity expected due to the highly conserved nature of Gia2. |
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| 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. |
