| Catalog # | A1059-77 |
| Applications | Suitable for use in Western Blot. Other applications have not been tested. |
| Recommended Dilution | Western Blot Analysis: 1ug/ml detects AIF from a mouse liver preparation. Mouse liver protein preparation was resolved by electrophoresis, transferred to nitrocellulose and probed with AIF (1ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. |
| Optimal dilutions to be determined by the researcher. |
| Western Blot Protocol 1. Perform SDS-PAGE on a sample and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water. 2. Block the blotted nitrocellulose in freshly prepared TBS containing 3% nonfat dry milk with 0.1% Tween-20 (TBST-MLK) for 1 hour at RT with constant agitation. 3. Incubate the nitrocellulose with 1ug/ml of AIF, A1059-77 diluted in freshly prepared TBS-MLK for three hours with agitation at RT 4. Wash the nitrocellulose twice with water. 5. Incubate the nitrocellulose in the secondary reagent of choice (1 | 3000 dilution was used) in TBS-MLK overnight at 4ºC with agitation. |
| 6. Wash the nitrocellulose with water three times. |
| 7. Wash the nitrocellulose in TBS-0.1% Tween 20 for 30 minutes with agitation at RT. |
| 8. Rinse the nitrocellulose in water for 30 minutes with agitation at RT. |
| 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 | Polyclonal |
| Host | Rabbit |
| Source | Human |
| Concentration | ~1mg/ml |
| Form | 200ug of protein A purified rabbit IgG in 200ul of 0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%. Liquid at -20°C. |
| Purity | Purified by Protein A affinity chromatography. |
| Immunogen | KLH-conjugated, synthetic peptide corresponding to amino acids 593–606 of human Apoptosis Inducing Factor, AIF (C-KDGEQHEDLNEV-AK), containing a N-terminal cysteine. The immunizing sequence is identical in mouse and rat. |
| Specificity | Recognizes AIF, Mr 57kDa. Another non-specific protein was also detected at Mr 120kDa. Species Crossreactivity: Mouse. Predicted for human and rat based on sequence homology |
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| Important Note | This product is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
