RPL37 | Ribosomal protein L37 (cytoplasmic)
AS12 2115 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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Recombinant, full length RPL37 of Chlamydomonas reinhardtii, UniProt: A8IBG1, 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 1:10 000 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.
Courtesy of Dr. Silvia Ramundo, University of Geneva, Switzerland
Reactant: Chlamydomonas reinhardtii (Green Alga)
Application: Western Blotting
Pudmed ID: 29053817
Journal: J Exp Bot
Figure Number: 3A,B,C,D
Published Date: 2018-03-14
First Author: Couso, I., Pérez-Pérez, M. E., et al.
Impact Factor: 6.088Open Publication
Inhibition of autophagic flux by concanamycin A (ConcA) prevents the degradation of ribosomal proteins under nitrogen starvation or phosphate limitation. (A) Chlamydomonas cells growing exponentially in Tris-acetate phosphate medium (TAP) were washed twice with nitrogen-free medium (TAP-N) and grown under these conditions for 4, 8, and 24 h. Control cells were washed in TAP medium and grown in the presence of nitrogen. (B) Chlamydomonas cells growing in TAP medium were washed with a nitrogen-free medium and grown under these conditions for 16 h in the absence (–) or presence (+) of 0.1 µM ConcA. (C) Chlamydomonas cells growing exponentially in TAP medium were washed with a phosphate-free medium (TA) and grown under these conditions during 8, 24, and 48 h. Control cells were washed and resuspended in TAP medium. (D) Chlamydomonas cells growing in TAP medium were washed with a phosphate-free (TA) medium and grown under these conditions for 48 h. Before collecting samples, cells were treated for 24 h with 0.1 µM ConcA. For (A–D), 20 µg of total extracts were resolved by 12% (RPS6) or 15% (RPL37, ATG8, and FKBP12) SDS–PAGE followed by western blotting with anti-OLLAS, anti-RPL37, anti-ATG8, and anti-FKBP12 antibodies. Molecular mass markers (kDa) are indicated on the left.
RPL37 (Ribosomal protein L37) belongs to the ribosomal protein L37e family (cytosolic large ribosomal subunit) and binds to the 23S rRNA.
Gonzaga Heredia-Martinez et al. (2018). Chloroplast damage induced by the inhibition of fatty acid synthesis triggers autophagy in Chlamydomonas. Plant Physiol, Sept. 2018.
Couse et al. (2017). Autophagic flux is required for the synthesis of triacylglycerols and ribosomal protein turnover in Chlamydomonas. J Exp Bot. 2017 Oct 19. doi: 10.1093/jxb/erx372.
Couso et al. (2017). Autophagic flux is required for the synthesis of triacylglycerols and ribosomal protein turnover in Chlamydomonas. J Exp Bot. 2017 Oct 19. doi: 10.1093/jxb/erx372.
Ramundo et al. (2013). Repression of Essential Chloroplast Genes Reveals New Signaling Pathways and Regulatory Feedback Loops in Chlamydomonas. The Plant Cell.
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