AIF Antibody

AIF Antibody: Apoptosis is characterized by several morphological nuclear changes including chromatin condensation and nuclear fragmentation. These changes are triggered by the activation of members of caspase family, caspase activated DNase, and several novel proteins. A novel gene, the product of which causes chromatin condensation and DNA fragmentation, was recently identified, cloned, and designated apoptosis inducing factor (AIF) . Like the critical molecules, cytochrome c and caspase-9, in apoptosis, AIF localizes in mitochondria. AIF translocates to the nucleus when apoptosis is induced and induces mitochondria to release the apoptogenic proteins cytochrome c and caspase-9. AIF induces chromatin condensation and large scale DNA fragmentation, which are the hallmarks of apoptosis, of the isolated nucleus and the nucleus in live cells by microinjection and apoptosis stimuli. AIF is highly conserved between human and mouse and widely expressed.

Independent Antibody Validation in Cell lines (Figure 2) shows similar AIF expression profile in human and mouse cell lines detected by three independent anti-AIF antibodies that recognize different epitopes, 2267 against central domain, 2239 against N-terminus domain and 2301 against C-terminus domain. AIF proteins are detected in the most tested cell lines at different expression levels by the three independent antibodies. Independent Antibody Validation in Cell lines2267 22392301 KD Validation (Figure 6,7) : Anti-AIF antibody specificity was further verified by AIF specific knockdown. AIF signal in H1299 cells and AIF stable cells transfected with AIF siRNAs was disrupted in comparison with that in cells transfected with control siRNAs. KD Validation Localization Validation (Figure 10) : AIF detected by anti-AIF antibodies is colocalized with Mitotracker (mitochondrion marker) . AIF is localized in mitochondria. Localization Validation Induced Expression Validation (Figure 11) : AIF expression detected by anit-AIF antibodies was up-regulated by Magnolol treatment. Induced Expression Validation

AIF antibody can be stored at 4˚ C for three months and -20˚ C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Human AIF has 6 isoforms, including isoform 1 (613aa, 67kD), isoform 2 (326aa, 36kD), isoform 3 (609aa, 66kD), isoform 4 (324aa, 35kD), isoform 5 (261aa, 28kD), and isoform 6 (237aa, 26kD) . Mouse AIF has one isoform (612aa, 67kD) and Rat AIF also has one isoform (612aa, 67kD) . 2267 can detect 4 human isoforms (1,2,3,5) and can also detect mouse and rat AIF.