For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human AIFM2/FSP1 protein. Antibodies are purified by peptide affinity chromatography.
Apoptosis-inducing factor mitochondria-associated 2 (AIFM2), also known as AIF-homologous mitochondrion-associated inducer of death (AMID) or p53-responsive gene 3 (PRG3), is a homolog of apoptosis-inducing factor (AIF) (1,2). Research studies show that it inhibits ferroptosis, a form of cell death caused by iron-dependent lipid peroxidation. Therefore, this protein was named as ferroptosis suppressor protein 1 (FSP1). FSP1 is a CoQ oxidoreductase and generates the reduced form of coenzyme Q10, which traps lipid peroxides, leading to the suppression of ferroptosis. Furthermore, inhibition of ferroptosis by FSP1 was shown to be independent of GPX4, another negative regulator of ferroptosis (3,4).
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