AIF Antibody Cat. No.: 2301

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psi-iconSpecifications
HOST SPECIES:Rabbit
SPECIES REACTIVITY: Human, Mouse, Rat
IMMUNOGEN: Anti-AIF antibody (2301) was raised against a peptide corresponding to 14 amino acids near the carboxy terminus of human AIF.

The immunogen is located within the last 50 amino acids of AIF.
TESTED APPLICATIONS: ELISA, ICC, WB
APPLICATIONS: WB: 0.5-2 μg/mL; ICC: 5 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunocytochemistry in human samples. All other applications and species not yet tested.
SPECIFICITY: Multiple isoforms of AIF are known to exist.
POSITIVE CONTROL:1) Cat. No. 1204 - K562 Cell Lysate
 2) Cat. No. 1205 - Jurkat Cell Lysate
 3) Cat. No. 17-005 - Jurkat Cell Slide
 4) Cat. No. 17-004 - K-562 Cell Slide
PREDICTED MOLECULAR WEIGHT: Predicted: 67kD

Observed: 68kD
psi-iconAdvanced Validation
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 9): AIF detected by anti-AIF antibodies is colocalized with Mitotracker (mitochondrion marker).  AIF is localized in mitochondria.

Induced Expression Validation (Figure 10): 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). 2301 can detect 4 human isoforms (1,2,3,5) and also can detect mouse and rat AIF.

psi-iconProperties
PURIFICATION:AIF Antibody is affinity chromatography purified via peptide column.
CLONALITY:Polyclonal
ISOTYPE:IgG
CONJUGATE:Unconjugated
PHYSICAL STATE:Liquid
BUFFER:AIF Antibody is supplied in PBS containing 0.02% sodium azide.
CONCENTRATION:batch dependent
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
OFFICIAL SYMBOL:CNKSR1
ALTERNATE NAMES:AIF Antibody: CNK, KSR, CNK1, Connector enhancer of kinase suppressor of ras 1, CNK homolog protein 1, Connector enhancer of KSR 1
ACCESSION NO.:O95381
PROTEIN GI NO.:50400606
GENE ID:10256
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 DNA fragmentation, which are the hallmarks of apoptosis, of the isolated nucleus and the nucleus in live cells by microinjection. AIF is highly conserved between human and mouse and widely expressed.
REFERENCES:1) Zamzami et al. Nature 1999; 401:127-8.
2) Susin SA, Lorenzo HK, Zamzami N, et al. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature 1999;397:441-6 (WD0800)
3) Susin et al. Nature 1999; 397:441-6.
4) Daugas et al. FASEB J. 2000; 14:729-39.
psi-iconCITATIONS
CITATIONS: 1)Stambolsky et al. Regulation of AIF expression by p53. Cell Death Differ. 2006;13(12):2140-9. Epub 2006. PMID: 16729031
2)Apostolova et al. Loss of apoptosis-inducing factor leads to an increase in reactive oxygen species, and an impairment of respiration that can be reversed by antioxidants. Cell Death Differ. 2006;13(2):354-7. PMID: 16195738
3)Hofer et al. Bacterial meningitis impairs hippocampal neurogenesis. J Neuropathol Exp Neurol. 2011;70(10):890-9. doi: 10.1097/NEN.0b013e3182303f31. PMID: 21937913
4)Gupta et al. Arsenic trioxide induces apoptosis in peripheral blood T lymphocyte subsets by inducing oxidative stress: a role of Bcl-2. Mol Cancer Ther. 2003;2(8):711-9. PMID: 12939460
5)Ikai et al. Magnolol-induced apoptosis is mediated via the intrinsic pathway with release of AIF from mitochondria in U937 cells. Biol Pharm Bull. 2006;29(12):2498-501. PMID: 17142989
6)Nebbioso et al. Tumor-selective action of HDAC inhibitors involves TRAIL induction in acute myeloid leukemia cells. Nat Med. 2005;11(1):77-84. Epub 2004. PMID: 15619633
7)Karlsson et al. Arsenic trioxide-induced death of neuroblastoma cells involves activation of Bax and does not require p53. Clin Cancer Res. 2004;10(9):3179-88. PMID: 15131059
8)Li et al. Apoptotic signaling pathways induced by nitric oxide in human lymphoblastoid cells expressing wild-type or mutant p53. Cancer Res. 2004;64(9):3022-9. PMID: 15126337
9)Cervera et al. Cells silenced for SDHB expression display characteristic features of the tumor phenotype. Cancer Res. 2008;68(11):4058-67. doi: 10.1158/0008-5472.CAN-07-5580. PMID: 18519664
10)Hahn et al. Galectin-1 induces nuclear translocation of endonuclease G in caspase- and cytochrome c-independent T cell death. Cell Death Differ. 2004;11(12):1277-86. PMID: 15297883
11)Hamacher-Brady et al. Response to myocardial ischemia/reperfusion injury involves Bnip3 and autophagy. Cell Death Differ. 2007;14(1):146-57. Epub 2006. PMID: 16645637
12)Nakagawa et al. Characterized mechanism of alpha-mangostin-induced cell death: caspase-independent apoptosis with release of endonuclease-G from mitochondria and increased miR-143 expression in human colorectal cancer DLD-1 cells. Bioorg Med Chem. 2007;15(16):5620-8. Epub 2007. PMID: 17553685
13)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. Epub 2007. PMID: 17980607
14)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
15)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. Epub 2005. PMID: 15699578
16)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
17)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
18)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
19)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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