AIF Antibody Cat. No.: 2267

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IMMUNOGEN: Anti-AIF antibody (2267) was raised against a peptide corresponding to 15 amino acids near the center of human AIF.

The immunogen is located within amino acids 500-550 of AIF.
APPLICATIONS: WB: 1-2 μg/mL; IHC-P: 10 μg/mL.

Antibody validated: Western Blot in human, mouse, and rat samples; Immunohistochemistry in human samples. All other applications and species not yet tested.
SPECIFICITY: At least five isoforms of AIF are known to exist; this antibody will detect all isoforms except isoform 5.
POSITIVE CONTROL:1) Cat. No. 1204 - K562 Cell Lysate
 2) Cat. No. 1461 - Rat Heart Tissue Lysate
 3) Cat. No. 1401 - Mouse Heart Tissue Lysate

Observed: 68 kDa
psi-iconAdvanced Validation

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. 

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.

Localization Validation (Figure 10): AIF detected by anti-AIF antibodies is colocalized with Mitotracker (mitochondrion marker).  AIF is localized in mitochondria.

Induced Expression Validation (Figure 11): AIF expression detected by anit-AIF antibodies was up-regulated by Magnolol treatment.

ISOFORMS: 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.

PURIFICATION:AIF Antibody is affinity chromatography purified via peptide column.
BUFFER:AIF Antibody is supplied in PBS containing 0.02% sodium azide.
STORAGE CONDITIONS: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.

psi-iconAdditional Info
ALTERNATE NAMES:AIF Antibody: Apoptosis-inducing factor 1, Programmed cell death protein 8, AIF, PDCD8
PROTEIN GI NO.:50400606
GENE ID:9131
USER NOTE:Optimal dilutions for each application to be determined by the researcher.
psi-iconBackground and References
BACKGROUND: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.
REFERENCES:1) Zamzami et al. Nature 1999; 401:127-8.
2) Susin et al. Nature 1999; 397:441-6
3) Daugas et al. FASEB J. 2000; 14:729-39
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14)Itoh T et al. Eupalinin A isolated from Eupatorium chinense L. induces autophagocytosis in human leukemia HL60 cells. Bioorg Med Chem. 2008;16(2):721-31.PMID: 17980607
15)Yel et al. Thimerosal induces neuronal cell apoptosis by causing cytochrome c and apoptosis-inducing factor release from mitochondria. Int J Mol Med. 2005;16(6):971-7. PMID: 16273274
16)Nakagawa et al. A potent apoptosis-inducing activity of a sesquiterpene lactone, arucanolide, in HL60 cells: a crucial role of apoptosis-inducing factor. J Pharmacol Sci. 2005;97(2):242-52. PMID: 15699578
17)Makani et al. Biochemical and molecular basis of thimerosal-induced apoptosis in T cells: a major role of mitochondrial pathway. Genes Immun. 2002;3(5):270-8. PMID: 12140745
18)Morse et al. Concurrent mRNA and protein extraction from the same experimental sample using a commercially available column-based RNA preparation kit. Biotechniques. 2006;40(1):54, 56, 58. PMID: 16454040
19)Itoh et al. Intracellular glutathione regulates sesquiterpene lactone-induced conversion of autophagy to apoptosis in human leukemia HL60 cells. Anticancer Res. 2009;29(4):1449-57. PMID: 19414401
20)Makani et al. Biochemical and molecular basis of thimerosal-induced apoptosis in T cells: a major role of mitochondrial pathway. Genes Immun. 2002;3(5):270-8. PMID: 12140745


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