RPN6 | 26S proteasome non-ATPase regulatory subunit 9
AS15 2832A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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15 µg or 10 µg or 5 µg of protein total extract from Arabidopsis thaliana are denatured at 70°C for 10 min then separated on 10% SDS-PAGE. Blotted 1 h 100V to Nitrocellulose membrane (GE Healthcare; 0.2 µm) tank transfert (Tris 50 mM; Glycine 40 mM; SDS 0.0375 % (m/v); methanol 20 % (v/v)). Blocked with TBS-T 0.1% powder milk 3% 1 h at room temperature. Blot α-RPN6 (AS15 2832A) was incubated 1 h at room temperature 1:1000 in TBS-T powder milk 3% and washed 2 times for 10 min with TBS T 0.1% at room temperature with agitation. Blot was incubated in secondary antibody anti-Rabbit IgG horse radish peroxidase conjugated diluted to 1:20 000 in TBS-T 0.1% from Agrisera (AS09 602) 1 h at room temperature and washed 3 times for 10 min with TBST 0.1%. Development with 20x LumiGLO ® Reagent and 20x Peroxide (#7003 cell signaling). Exposure time vary between 8 to 30 min.
Courtesy of Dr. Claire Rosnoblet (MCF), Agroécologie Agrosup Dijon / INRA / Université de Bourgogne, France
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. RPN6 associates with an ATPase subunit of the 19S proteasome regulatory complex, AtS6A. Alternative names: 26S proteasome non-ATPase regulatory subunit 11 homolog, 19S proteosome subunit 9, AtS9 26S proteasome regulatory subunit RPN6, AtRPN6, 26S proteasome regulatory subunit S9 homolog.