DS5a | Drosophila 26S proteasome subunit Rpn10
AS01 012 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Drosophila melanogaster
|Recommended dilution||1 : 2000 (WB)|
|Expected | apparent MW||
42.6 | 54 kDa (Drosophila), 40 kDa (A. thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Drosophila melanogaster|
|Predicted reactivity||Bovine, Mouse, Pig, Salmon, Rat,|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||Nguyen at al (2012). Anupstreamregulator of the26Sproteasomemodulatesorgansize in Arabidopsisthaliana. Plant J. Dec 17.
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.
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 (GE healthcare). Exposure time was 2 seconds. Position of protein on gel corresponds to expected size - 40 kDa.
Courtesy Dr. Jozefus Schippers, Max Planck Institute, Germany
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