L13-1 | 60S ribosomal protein L13-1
AS13 2650 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Hordeum vulgare
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|Recommended dilution||1 : 2500 (WB)|
|Expected | apparent MW||
23.7 | 29 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Hordeum vulgare|
|Predicted reactivity||Brassica napus, Chlamydomonas reinhardii, Nicotiana tabacum, Oryza sativa, Picea excelsa, Populus balsamifera, Sorghum bicolor, Ricinus communis, Zea mays, Vitis vinifera|
|Not reactive in||Diatoms|
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
to be added when available, antibody released in October 2013.
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 % RPN2125 (GE Healthcare) 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 TMA-6 (Lumigen) 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.
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