DS5a | Drosophila 26S proteasome subunit Rpn10
AS01 012 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Drosophila melanogaster
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Drosophila mealnogaster 26S Proteasome subunit S5a also known as Rpn10, p54. Antigen sequence is also conserved in Arabidopsis thaliana AT4G38630
42.6 | 54 kDa (Drosophila), 40 kDa (A. thaliana)
3 µg of total protein from Arabidopis thaliana leaves extracted with CelLytic P (Sigma) were separated on 10 % SDS-PAGE and blotted overnight at 30V to a nitrocellulose membrane. Blots were blocked with 1% western blocking reagent (Roche) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody (DS5a) at a dilution of 1:2000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly, and then washed twice for 10 min with TBS-T at RT with agitation. Subsequently, the blot was washed twice with 0,5% western blocking reagent for 10 min each. Blot was incubated in secondary antibody (Goat anti-rabbit IgG (H&L) HRP conjugated, AS09 602 from Agrisera) diluted to 1:50 000 in 0,5% western blocking reagent for 1h at RT with agitation. The blot was washed 4 times with TBS-T buffer. Detection of HRP was performed with ECL according to the manufacturer's instructions. Exposure time was 2 seconds. Position of protein on gel corresponds to expected size - 40 kDa.
Courtesy Dr. Jozefus Schippers, Max Planck Institute, Germany
Proteasome-dependent degradation serves an essential role in the removal of a wide variety of key nuclear and cytosolic proteins. Substrates are targeted for proteolysis by the ubiquitin pathway before being degraded by the 26 S proteasome. The subunit of the proteasome, S5a, was identified to bind to polyubiquitin in vitro and thus proposed to act as a substrate recognition component.
Lundgren et al. (2003). Use of RNA interference and complementation to study the function of the Drosophila and human 26S proteasome subunit S13. Mol Cel Biol 23:5320-5330.