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)
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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.
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