Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10% gel)
Loading amount
30 µg
Treatment
Fed, Fasted, Fasted on HFD
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 27°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% milk in PBSt
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: IRDye 800CW Donky Anti-Rabbit IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
LICOR
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 46 kDa Non-specific: 55, 49, 30 kDa
Positive control
HepG2 cell lysate (expressing human ACOT1)
Additional data
Additional Notes
mouse ACOT1 and ACOT2 are 93.7% homologous, however ACOT2 is localized to the mitochondria while ACOT1 is cytosolic. Our mouse ACOT1 antibody picks up both ACOT1 and ACOT2. The abcam100915 hACOT1 antibody did not recognize ACOT2 in addition to its lack of recognizing ACOT1 in a manner similar to what we see with our mouse ACOT1 antibody. ACOT1 is lowly expressed in the fed state, but is highly expressed in the fasted state.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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提交于 Nov 18 2016