Temporal Mitochondrial Proteome Changes

Mitochondrial samples, 4-11 weeks brain (n=5), 78 weeks brain (n=5), 4-11 weeks skeletal muscle (n=3) and 78 weeks skeletal muscle (n=3), were subject to iso-electric focussing using ZOOM IPG (Life Technologies) system and pH 3-10 (non-linear) ZOOM IPG strips following the manufacturers protocol. Gels were stained (SimplyBlue™ SafeStain, Life Technologies) and imaged (ImageQuant 300, GE Healthcare Life Sciences). Analysis was performed using SameSpots software (Totallab). Protein spots with a p value of less than 0.15 and a fold change greater than 1.2 were further analysed (one-way ANOVA). Proteins were identified from the gel pieces as described previously [5 (link)].

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Pollard A., Shephard F., Freed J., Liddell S, & Chakrabarti L. (2016). Mitochondrial proteomic profiling reveals increased carbonic anhydrase II in aging and neurodegeneration. Aging (Albany NY), 8(10), 2425-2434.

Publication 2016

ZOOM IPG ZOOM IPG strips SimplyBlue™ SafeStain ImageQuant 300

Anova Brain Electric Mitochondrial Proteins Skeletal muscle Spots

Corresponding Organization : University of Nottingham

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Variable analysis

independent variables

Age of brain samples (4-11 weeks, 78 weeks)
Tissue type (brain, skeletal muscle)

dependent variables

Protein expression profiles (identified from gel pieces)

control variables

Experimental protocol (iso-electric focusing using ZOOM IPG system and pH 3-10 (non-linear) ZOOM IPG strips)
Staining (SimplyBlue™ SafeStain)
Imaging (ImageQuant 300, GE Healthcare Life Sciences)
Analysis software (SameSpots)

controls

No positive or negative controls were explicitly mentioned.

Annotations

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