| application information | ||
| Recommended dilution | 1: 2500 with standard ECL (WB) | |
| Expected | apparent MW | 23.7 | 29 kDa (Arabidopsis thaliana) | |
| Confirmed reactivity | Arabidopsis thaliana, Hordeum vulgare | |
| Predicted reactivity | dicots including: Brassica napus, Nicotiana tabacum, Ricinus communis, Vitis vinifera, monocots including: Oryza sativa, Sorghum bicolor, Zea mays, trees: Picea excelsa, Populus balsamifera, algae: Chlamydomonas reinhardii, | |
| Not reactive in | diatoms | |
| Additional information | Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel. | |
| Selected references | to be added when available, antibody released in October 2013. |
![\"western](\"https://www.ebiomall.com/images/agrisera/201709/AS13_2650.jpg\" "\\"western")
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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