RPS12 | Ribosomal protein S12 (chloroplastic)
AS12 2114 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Chlamydomonas reinhardtii
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Recombinant full length RPS12 of Chlamydomonas reinhardtii, P14149, expressed in E.coli
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10 µg of total protein from Chlamydomonas reinhardtii extracted with standard lysis buffer (Tris-HCl pH 6.8 50mM, 10mM EDTA, 2% SDS, Sigma protease inhibitors) were separated on 15 % SDS-PAGE and blotted to nitrocellulose membrane using a wet transfer cell. Blot was blocked in TBS-T containing 5% non-fat dry milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 in TBS-T containing 5% non-fat dry milk for 1h at RT with agitation. The antibody solution was decanted and the blot was washed 3 times for 15 min in TBS-T containing 1% non-fat dry milk with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:0 000 in TBS-T containing 1% non-fat dry milk for 1h at RT with agitation. The blot was washed as above and developed for 1 min with ECL according to the manufacturers instructions. Exposure time was 30 seconds.
RPS12 (ribosomal protein S12) is a plastid ribosomal protein which is a part of the 30S ribosomal subunit. Together with S4 and S5 plays an important role in translational accuracy.
Zoschke et al. (2016). The PPR-SMR protein PPR53 enhances the stability and translation of specific chloroplast RNAs in maize. Plant J. 2016 Mar;85(5):594-606. doi: 10.1111/tpj.13093. Epub 2016 Feb 5.
Ramundo et al. (2013). Repression of Essential Chloroplast Genes Reveals New Signaling Pathways and Regulatory Feedback Loops in Chlamydomonas. The Plant Cell.