Product Info

Source:
Mouse
Application:
ELISA 1:10000, WB 1:500-1:2000, IHC 1:200-1:1000, IF/ICC 1:200-1:1000, FCM 1:200-1:400
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat,Monkey
Clonality:
Monoclonal [AFB1912]
Specificity:
AIF antibody detects endogenous levels of total AIF.
RRID:
AB_2833935
Cite Format: Affinity Biosciences Cat# BF0591, RRID:AB_2833935.
Conjugate:
Unconjugated.
Purification:
Affinity-chromatography.
Storage:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AIFM1; AIFM1_HUMAN; Apoptosis inducing factor 1, mitochondrial; Apoptosis inducing factor; Apoptosis inducing factor, mitochondrion associated, 1; Apoptosis-inducing factor 1; CMTX4; COWCK; COXPD6; Harlequin; Hq; mAIF; MGC111425; MGC5706; mitochondrial; Neuropathy, axonal motor-sensory, with deafness and mental retardation; neuropathy, axonal, motor-sensory with deafness and mental retardation (Cowchock syndrome); PDCD 8; PDCD8; Programmed cell death 8 (apoptosis inducing factor); Programmed cell death 8; Programmed cell death 8 isoform 1; Programmed cell death 8 isoform 2; Programmed cell death 8 isoform 3; Programmed cell death protein 8; Programmed cell death protein 8 mitochondrial; Programmed cell death protein 8 mitochondrial precursor; Programmed cell death protein 8 mitochondrial precursor; Striatal apoptosis inducing factor;

Immunogens

Immunogen:

Purified recombinant fragment of human AIF expressed in E. Coli.

Uniprot:
Gene(ID):
Expression:
O95831 AIFM1_HUMAN:

Expressed in all tested tissues (PubMed:16644725). Detected in muscle and skin fibroblasts (at protein level) (PubMed:23217327).

Brain specific.

Expressed in all tested tissues except brain.

Isoform 5 is frequently down-regulated in human cancers.

Description:
This gene encodes a flavoprotein essential for nuclear disassembly in apoptotic cells, and it is found in the mitochondrial intermembrane space in healthy cells. Induction of apoptosis results in the translocation of this protein to the nucleus where it affects chromosome condensation and fragmentation. In addition, this gene product induces mitochondria to release the apoptogenic proteins cytochrome c and caspase-9. Mutations in this gene cause combined oxidative phosphorylation deficiency 6, which results in a severe mitochondrial encephalomyopathy. Alternative splicing results in multiple transcript variants. A related pseudogene has been identified on chromosome 10
Sequence:
MFRCGGLAAGALKQKLVPLVRTVCVRSPRQRNRLPGNLFQRWHVPLELQMTRQMASSGASGGKIDNSVLVLIVGLSTVGAGAYAYKTMKEDEKRYNERISGLGLTPEQKQKKAALSASEGEEVPQDKAPSHVPFLLIGGGTAAFAAARSIRARDPGARVLIVSEDPELPYMRPPLSKELWFSDDPNVTKTLRFKQWNGKERSIYFQPPSFYVSAQDLPHIENGGVAVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTLFRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILPEYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEPNVELAKTGGLEIDSDFGGFRVNAELQARSNIWVAGDAACFYDIKLGRRRVEHHDHAVVSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQDNPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGIVLWNIFNRMPIARKIIKDGEQHEDLNEVAKLFNIHED

PTMs - O95831 As Substrate

Site PTM Type Enzyme
S56 Phosphorylation
S57 Phosphorylation
S60 Phosphorylation
T87 Phosphorylation
Y95 Phosphorylation
S100 Phosphorylation
T105 Phosphorylation
K109 Ubiquitination
K111 Ubiquitination
S116 Phosphorylation
S118 Phosphorylation
S176 Phosphorylation
S182 Phosphorylation
T188 Phosphorylation
K189 Ubiquitination
T190 Phosphorylation
K232 Ubiquitination
K244 Acetylation
K255 Ubiquitination
T263 Phosphorylation
S266 Phosphorylation
S268 Phosphorylation
K278 Ubiquitination
S279 Phosphorylation
S292 Phosphorylation
K295 Acetylation
T328 Phosphorylation
K337 Acetylation
Y347 Phosphorylation
S371 Phosphorylation
S375 Phosphorylation
S376 Phosphorylation
S416 Phosphorylation
Y443 Phosphorylation
K518 Ubiquitination
S519 Phosphorylation
T521 Phosphorylation
S524 Phosphorylation
T526 Phosphorylation
S530 Phosphorylation
S532 Phosphorylation
T542 Phosphorylation
S546 Phosphorylation
T547 Phosphorylation
R569 Methylation
K593 Acetylation
K593 Ubiquitination
K606 Ubiquitination

Research Backgrounds

Function:

Functions both as NADH oxidoreductase and as regulator of apoptosis. In response to apoptotic stimuli, it is released from the mitochondrion intermembrane space into the cytosol and to the nucleus, where it functions as a proapoptotic factor in a caspase-independent pathway. The soluble form (AIFsol) found in the nucleus induces 'parthanatos' i.e. caspase-independent fragmentation of chromosomal DNA (By similarity). Binds to DNA in a sequence-independent manner. Interacts with EIF3G, and thereby inhibits the EIF3 machinery and protein synthesis, and activates caspase-7 to amplify apoptosis. Plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells. In contrast, participates in normal mitochondrial metabolism. Plays an important role in the regulation of respiratory chain biogenesis by interacting with CHCHD4 and controlling CHCHD4 mitochondrial import.

Has NADH oxidoreductase activity. Does not induce nuclear apoptosis.

Pro-apoptotic isoform.

PTMs:

Under normal conditions, a 54-residue N-terminal segment is first proteolytically removed during or just after translocation into the mitochondrial intermembrane space (IMS) by the mitochondrial processing peptidase (MPP) to form the inner-membrane-anchored mature form (AIFmit). During apoptosis, it is further proteolytically processed at amino-acid position 101 leading to the generation of the mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis in a caspase-independent manner.

Ubiquitination by XIAP/BIRC4 does not lead to proteasomal degradation. Ubiquitination at Lys-255 by XIAP/BIRC4 blocks its ability to bind DNA and induce chromatin degradation, thereby inhibiting its ability to induce cell death.

Subcellular Location:

Mitochondrion intermembrane space. Mitochondrion inner membrane. Cytoplasm. Nucleus. Cytoplasm>Perinuclear region.
Note: Proteolytic cleavage during or just after translocation into the mitochondrial intermembrane space (IMS) results in the formation of an inner-membrane-anchored mature form (AIFmit). During apoptosis, further proteolytic processing leads to a mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis (PubMed:15775970). Colocalizes with EIF3G in the nucleus and perinuclear region (PubMed:17094969).

Mitochondrion intermembrane space. Mitochondrion inner membrane.
Note: Has a stronger membrane anchorage than isoform 1.

Mitochondrion. Cytoplasm>Cytosol.
Note: In pro-apoptotic conditions, is released from mitochondria to cytosol in a calpain/cathepsin-dependent manner.

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in all tested tissues. Detected in muscle and skin fibroblasts (at protein level).

Brain specific.

Expressed in all tested tissues except brain.

Isoform 5 is frequently down-regulated in human cancers.

Subunit Structure:

Monomer (oxidized form). Homodimer (reduced form). Upon reduction with NADH, undergoes dimerization and forms tight, long-lived FADH2-NAD charge transfer complexes (CTC) resistant to oxidation. Also dimerizes with isoform 3 preventing its release from mitochondria. Interacts with XIAP/BIRC4. Interacts (via N-terminus) with EIF3G (via C-terminus). Interacts with PRELID1. Interacts with CHCHD4; the interaction increases in presence of NADH.

Family&Domains:

Belongs to the FAD-dependent oxidoreductase family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

References

1). Zeng M et al. Danhong injection alleviates cerebral ischemia/reperfusion injury by improving intracellular energy metabolism coupling in the ischemic penumbra. Biomedicine & Pharmacotherapy 2021 Aug;140:111771. (PubMed: 34058441) [IF=7.5]

Application: WB    Species: rat    Sample:

Fig. 4.| Effect of DHI on the activity of PARP1/AIF signaling pathway and the content of molecules associated with cytoplasmic glycolysis. A-E: Representative images of WB analysis and the semi-quantification of PARP1, PAR, AIF, and HSP70. Data are expressed as mean ± SD (n = 3).

Application: IHC    Species: rat    Sample: brain

Fig. 5.| Effect of DHI on the content of molecules associated with cytoplasmic glycolysis. A-F: Immunostaining photomicrographs of PARP1, AIF, HSP70 and quantitative analysis of the IOD.

2). Li et al. Mesenchymal stem cell‑derived extracellular vesicles prevent neural stem cell hypoxia injury via promoting miR‑210‑3p expression. Molecular Medicine Reports 2020 Nov;22(5):3813-3821. (PubMed: 33000190) [IF=3.4]

Application: WB    Species: rat    Sample: NSCs

Figure 6. |miR‑210-3p inhibitor promotes hypoxia injury-induced AIF and BNIP3 expression in NSCs. miR‑210-3p expression levels were determined by western blotting and quantified. *P<0.05 vs. control; #P<0.05 vs. model(n=5 per group). miR, microRNA; AIF, apoptosis-inducing factor; BNIP3,Bcl-2 19 kDa interacting protein.

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