Santa Cruz
AIF (clone E1, SantaCruz; 1/1000).
sc-13116
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HEK293 cells were fractionated into cytosol and heavy membrane fractions. AIF was utilized to track the purity of the fractionation as AIF is a mitochondrial protein.
Western Blot
HEK293 cell lysates- fractionation into cytosol and Heavy membrane
5% skim milk in TBST with anti-AIF 1/1000- 1 hour at room temperature
5% skim milk in TBST- 10 minutes at room temperature
1/10000, RT for 1 hour in TBST 5% skim milk
NA
HRP- chemiluminescence
AIF was detected as expected in the total (T) and heavy membrane (HM) fractions as expected, suggesting that the fractionation was fairly clean. Lanes 1-3 (control) and Lanes 4-6 (stimulated) represent separate samples that have been fractionated as shown.
A good antibody for mitochondrial staining.
None
A good mitochondrial marker that requires a low amount of antibody for detection.