PR-1 | Pathogenesis-related protein 1
AS10 687 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
|Recommended dilution||1 : 2500 (WB)|
|Expected | apparent MW||
17.7 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Hordeum vulgare, Nicotiana bentamiana, Spinacia oleracea, Solanum lycopersicum, Triticum aestivum, Zea mays|
Brassica rapa subsp. pekinensis, Brassica napus, Eutrema japonica, Glycine max, Phaseolus vulgaris, Solanum tuberosum
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
Re-using of antibody solution is not recommended. It will contribute to incrteased background signal.
|Selected references||Izquierdo et al. (2018). Arabidopsis nonresponding to oxylipins locus NOXY7 encodes a yeast GCN1 homolog that mediates noncanonical translation regulation and stress adaptation. Plant Cell Environ. 2018 Mar 2. doi: 10.1111/pce.13182.
Seguel et al. (2018). PROHIBITIN 3 forms complexes with ISOCHORISMATE SYNTHASE 1 to regulate stress-induced salicylic acid biosynthesis in Arabidopsis. Plant Physiol. Jan 2018. DOI:10.1104/pp.17.00941
Huh et al. (2017). Protein-protein interactions in the RPS4/RRS1 immune receptor complex. PLoS Pathog. 2017 May 5;13(5):e1006376. doi: 10.1371/journal.ppat.1006376.
Zhang et al. (2017). A suite of receptor-like kinases and a putative mechano-sensitive channel are involved in autoimmunity and plasma membrane-based defenses in Arabidopsis. Mol Plant Microbe Interact. 2017 Jan 4. doi: 10.1094/MPMI-09-16-0184-R.
Zhu et al. (2016). CML8, an Arabidopsis calmodulin-like protein plays a role in Pseudomonas syringae plant immunity. Plant Cell Physiol. 2016 Nov 10. pii: pcw189. [Epub ahead of print]
Recombinant PR-1 protein standard 0.05 pmol (1), 0.1 pmol (2), 0.15 pmol (3), 0.2 pmol (4), and 0.3 pmol (5) was loaded in each lane. 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 anti-PR-1 antibody at a dilution of 1: 10 000 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 1 minute.
Immunostaining of Arabidopsis thaliana seedlings treated with 250 µM SA for 120 minutes for induction of PR- gene expression (right panel) or control without SA (left panel). Steps involved: fixation: in 2 % formaldehyde in MTSB buffer for 30 minutes at 37 °C; washing with water, hydrophilization with methanol; cell wall digestion: 5-7 minutes, 0.25% Dricelaze, 0.1 % Macerozyme in 5 mM MES buffer, pH5.2; cell wall permeabilization: 10 % DMSO/3 % NP40 in MTSB buffer; primary antibody incubation: dilution 1: 200 in MTBS buffer for 3 h at RT; secondary antibody incubation: dilution 1: 1000 at RT for 1 h, goat-anti rabbit IgG Alexa 488 conjugated antibody.
Courtesy of Dr. Taras Pasternak, Freiburg University, Germany
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