RPS12 | Ribosomal protein S12 (chloroplastic)
AS12 2114 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Chlamydomonas reinhardtii
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|Recommended dilution||1 : 10 000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana, Chlamydomonas reinhardtii, Zea mays|
Bacteria, Dunaliella salina, Chlorella vulgaris, Oltmannsiellopsis viridis (marine flagellate), Pinus sp., Physcomitrella patens, Prochlorococcus marinus, Scenedesmus obliquus, Spinacia oleracea, Synechocystis sp. PCC6803, Synechococcus elongatus PCC 7942, Volvox carteri (green alga)
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||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.
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.
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