Sensitive Quantitative Fluorescence Detection of Proteins in Gels Using SYPRO® Ruby Protein Gel Stain

Schulenberg, Birte; Ahnert, Nancy; Patton, Wayne F.

doi:10.1385/1-59259-890-0:209

Cited by 2 publications

(1 citation statement)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fluorescent stains like Sypro Ruby have a broad linear dynamic range (Nishihara and Champion, 2002) and are frequently used as loading controls and for quantitative protein determination following gel electrophoresis and western blotting, as demonstrated previously (Schulenberg et al, 2005;Aldridge et al, 2008;Lush et al, 2008;David et al, 2010). Hence, in line with such an approach, intensity values of MDA and Ub protein adducts were corrected for background staining and normalised to the total protein load within the 14-200kDa gel as detected by Sypro Ruby total protein staining following western blotting.…”

Section: Detection Of Mda and Ub Adducts In Brain Protein Fractionsmentioning

confidence: 99%

Flight restriction prevents associative learning deficits but not changes in brain protein-adduct formation during honeybee ageing

Tolfsen

Baker

Kreibich

et al. 2011

Journal of Experimental Biology

View full text Add to dashboard Cite

SUMMARYHoneybees (Apis mellifera) senesce within 2weeks after they discontinue nest tasks in favour of foraging. Foraging involves metabolically demanding flight, which in houseflies (Musca domestica) and fruit flies (Drosophila melanogaster) is associated with markers of ageing such as increased mortality and accumulation of oxidative damage. The role of flight in honeybee ageing is incompletely understood. We assessed relationships between honeybee flight activity and ageing by simulating rain that confined foragers to their colonies most of the day. After 15days on average, flight-restricted foragers were compared with bees with normal (free) flight: one group that foraged for ~15days and two additional control groups, for flight duration and chronological age, that foraged for ~5days. Free flight over 15days on average resulted in impaired associative learning ability. In contrast, flight-restricted foragers did as well in learning as bees that foraged for 5days on average. This negative effect of flight activity was not influenced by chronological age or gustatory responsiveness, a measure of the bees' motivation to learn. Contrasting their intact learning ability, flight-restricted bees accrued the most oxidative brain damage as indicated by malondialdehyde protein adduct levels in crude cytosolic fractions. Concentrations of mono-and poly-ubiquitinated brain proteins were equal between the groups, whereas differences in total protein amounts suggested changes in brain protein metabolism connected to forager age, but not flight. We propose that intense flight is causal to brain deficits in aged bees, and that oxidative protein damage is unlikely to be the underlying mechanism.Supplementary material available online at

show abstract