NPR1 Nonexpresser of PR genes 1 (1 mg)
AS12 1854-1mg Clonality: Polyclonal Host: Rabbit | Reactivity: Arabidopsis thaliana

Info: | Add review |
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application information |
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Recommended dilution | 1 : 1000 (WB) | |
Expected | apparent MW | 66 | 66 kDa |
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Confirmed reactivity | Arabidopsis thaliana | |
Predicted reactivity | Arabidopsis thaliana | |
Not reactive in | Nicotiana benthamiana, Nicotiana tabacum, Solanum lycopersicum, Solanum tuberosum |
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Additional information | Please note that depending upon detection system you are using, longer exposure time may be required with this antibody. |
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Selected references | to be added when available, antibody released in December 2015. |
application example

Samples (0.2 g) were collected from leaf tissue (3 week old rosettes; 24 hours after +/- 0.5 mM sodium salicylate spray). Total protein was extracted in a buffer containing 50 mM Tris-Hcl, pH 7.5; 150 mM NaC; 0.5 mM EDTA; 0.1 % Triton X-100; 0.2 % nonidet P-40; 50 uM MG115 and samples were adjusted to equal total protein concentration. Samples were denatured with 4X SDS Sample buffer with 200 mM DTT (final sample concentration of 50 mM) at 75 C for 15 min. Protein samples (30 µg of total protein) were separated on 4-12% Bis-Tris gel and blotted to PVDF using an iBlot (semi-dry transfer system; Life technologies). The membrane was blocked with 1X PBS-T containing 5% low fat dry milk ans 0.1 % tween-20, for 1h at room temperature (RT) with agitation and incubated in the primary antibody (in blocking solution) at a dilution of 1: 1 000 over night at 4 C with agitation. The antibody solution was decanted and the membrane was washed with blocking solution, 3 times for 10 min each at RT with agitation. The membrane was then incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in blocking solution for 1h at RT with agitation. The membrane was washed as described above, rinsed with 1x PBS-T and developed for 2 min with Pierce West Dura extended duration substrate. Exposure time was 30 seconds.
** Note: 50 mM DTT concentration in the extraction buffer is necessary to reduce NPR1 oligomer formation.
** Note: The expression level of endogenous NPR1 is very low in extracts from healthy, untreated plants. A successful SA treatment or pathogen infection is absolutely necessary for good detection.
Courtesy Dr. John Withers, Duke University, USA
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