兔抗拟南芥致病相关蛋白1（PR-1）多克隆抗体

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兔抗拟南芥致病相关蛋白1（PR-1）多克隆抗体介绍：

货   号：AS10 687

中文名称：兔抗拟南芥致病相关蛋白1（PR-1）多克隆抗体

英文名称：PR-1|Pathogenesis-related protein 1

应用：western blot (WB)

规格：50 µl

价格：4440元

兔抗拟南芥致病相关蛋白1（PR-1）多克隆抗体简介：

Application example

western blot -1 


 

Recombinant PR-1 protein (0.2 pmol) (7) and 20 µg of a total protein from Arabidopsis thaliana (1), Hordeum vulgare  (2), Zea mays (3),Spinacia oleracea (4), Solanum esculentum (5), Triticum aestivum (6) were extracted with Protein Extraction Buffer PEB (AS08 300),  recombinant PR-1 (7) and were supplemented with 50 mM DTT and heat at 70°C for 5 min and kept 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: 2 500 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 1:15 000 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 1 minute.

western blot -2

10 µg of total protein from Arabidopsis thaliana columbia strain from various experimental set ups, extracted with Sigma Aldrich Plant Total Extraction Kit were separated on 15 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 for 1h at RT 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 0.1% at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from BioRad) diluted to 1:3 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Biorad Opti-4Cn Substrate kit according to the manufacturer's instructions.

immunolocalization