G Protein, i alpha 2, (Gia2)

Applications: \nSuitable for use in ELISA and Immunoblot. Other applications not tested.\n\nRecommended Dilution: \nImmunoblot 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.\nOptimal dilutions to be determined by the researcher.\n\nImmunoblot Protocol: \n1. Perform SDS-PAGE on a microsomal membrane preparation. Transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. \n2. Block the blotted nitrocellulose in freshly prepared PBS, 3% nonfat dry milk (PBS-MLK) for 60 minutes at RT with constant agitation. \n3. Incubate the nitrocellulose with 1-2ug/ml of G8599-09, diluted in freshly prepared PBS-MLK, overnight at 4 degrees C with constant agitation.\n4. Wash the nitrocellulose twice with water. \n5. 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. \n6. Wash the nitrocellulose with water twice. \n7. Wash the nitrocellulose in PBS, 0.05% Tween-20 for 10 minutes. \n8. Rinse the nitrocellulose in 4-5 changes of water for 2 hours. \n9. Use detection method of choice. Enhanced chemiluminescence was used.\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees 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.

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