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Technical Data Sheet

AX5505

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phospho-AIFM1 (Ser-116)
Blocking Peptide
Price
Size

$55
50 μg

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Application
Dilution


Blocking
1:1000


ELISA
50 ng/well



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Apoptosis-inducing factor (AIFM1, AIF, PDCD8) is a ubiquitously expressed flavoprotein that plays a critical role in caspase-independent apoptosis. AIFM1 is expressed as a 66 kDa precursor protein before being N-terminally cleaved to 62 kDa and localized to the mitochondrial intermembrane space. In response to apoptotic stimuli, AIFM1 is released from the mitochondrial intermembrane as a 57 kDa fragment that can translocate to the nucleus. Treatment of isolated nuclei with recombinant AIFM1 leads to early apoptotic events, such as chromatin condensation and large-scale DNA fragmentation. Studies of AIFM1 knockout mice have shown that the apoptotic activity of AIFM1 is cell type and stimuli-dependent. Also, AIFM1 was required for embryoid body cavitation, representing the first wave of programmed cell death during embryonic morphogenesis. Structural analysis of AIFM1 revealed two regions, an oxidoreductase activity and a DNA binding domain. AIFM1 has dual functions, a pro-apoptotic activity in the nucleus via its DNA binding and an anti-apoptotic activity via the scavenging of free radicals through its oxidoreductase activity.


Background References
Daugas, E. et al. (2000) FEBS Lett. 476:118.
Lipton, S.A. and Bossy-Wetzel, E. (2002) Cell 111:147.
Delavallee, L. et al. (2011) IUBMB Life. 63(4):221.
Sequence
Phospho-AIFM1 (Ser-116) synthetic peptide corresponding to amino acids surrounding Ser-116 in human AIFM1. This site is conserved in rat and mouse AIFM1, but the amino acids surrounding the site are not well conserved. The site is not found in AIFM2 and AIFM3.
Buffer and Storage
Blocking Peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide.
Store at –20°C. Stable for 1 year.
Specificity
The peptide is specifically recognized by AIFM1 (Ser-116) phospho-specific antibody (AP5501) in ELISA, and has been shown to block the reactivity of AP5501 in Western blot. In addition, the peptide is recommended for use in blocking AP5501 reactivity in immunocytochemistry.
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