L13-1 | 60S ribosomal protein L13-1
AS13 2650 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Hordeum vulgare
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23.7 | 29 kDa (Arabidopsis thaliana)
10 µg of total protein from Arabidopsis thaliana (1) and Hordeum vulgare (2) leaf, extracted with Protein Extraction Buffer PEB (AS08 300), were supplemented with 50 mM DTT and heat at 70°C for 5 min and keep on ice before loading. Protein separation was done using NuPage 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2-2.5 % blocking reagent, in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 2500 in blocking reagent for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:25 000 in TBS-T for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 2 minutes.
Note: western blot detection pattern can be different if other type of samples is used, for example membrane fraction.
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
L13 protein is localized in the cytoplasm. It belongs to the ribosomal protein L13e family. Alternative name: protein BBC1 homolog.
Li (2019). The Isolation of Total and Membrane-Bound Polysomes from Arabidopsis and the Detection of Their Associated AGO1 and sRNAs. Methods Mol Biol. 2019;1932:317-333. doi: 10.1007/978-1-4939-9042-9_23.
You et al. (2019). FIERY1 promotes microRNA accumulation by suppressing rRNA-derived small interfering RNAs in Arabidopsis. Nat Commun. 2019 Sep 27;10(1):4424. doi: 10.1038/s41467-019-12379-z.
de Francisco Amorim et al. (2018). The U1 snRNP Subunit LUC7 Modulates Plant Development and Stress Responses via Regulation of Alternative Splicing. Plant Cell. 2018 Nov;30(11):2838-2854. doi: 10.1105/tpc.18.00244. Epub 2018 Oct 11.
Beine-Golovchuk et al. (2018). Plant Temperature Acclimation and Growth Rely on Cytosolic Ribosome Biogenesis Factor Homologs. Plant Physiol. 2018 Mar;176(3):2251-2276. doi: 10.1104/pp.17.01448. Epub 2018 Jan 30.
Wang et al. (2016). Comprehensive proteomic analysis of developing protein bodies in maize (Zea mays) endosperm provides novel insights into its biogenesis. J Exp Bot. 2016 Dec;67(22):6323-6335. Epub 2016 Oct 27.
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