PR-5 | Pathogenesis-related protein 5 (A.thaliana)
AS12 2373 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
|Info:||More information||Product suggestions||Add review|
1 : 10 000 (WB)
|Expected | apparent MW||Propeptide 25.3 kD, processing aa 1-23, mature peptide 22.8 kD|
|Confirmed reactivity||Arabidopsis thaliana
|Predicted reactivity||Arabidopsis thaliana, for other species check product AS15 2858|
|Not reactive in|
To be added when available, antibody released in January 2018.
50 and 100 µg of total protein from Arabidopsis thaliana wild-type plants treated with water and salicylic acid (SA) analog BTH (Benzothiadiazole) were extracted with buffer containing 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10 mM MgCl2, 5 mM EDTA, 10% glycerol, 0.1% NP-40, 0.1% protease inhibitor cocktail . The proteins were denatured by boiling for 5 min were separated on 10% SDS-PAGE and blotted 30 min to PVDF using tank transfer. Blots were blocked with TBST containing 5% non-fat dry milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 for for 4 h/RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 15 min each in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed with Clarity MAX (Bio-RAD). Exposure time was 3 minutes.
Courtesy Ms. Ruiying Liu and Dr. Pradeep Kachroo, University of Kentucky, USA
Arabidopsis thaliana Col-0 treated with water (1) Col-0 treated with 0.5 mM SA (2), npr1-2 treated with water (3), npr1-2 treated with 0.5 mM SA (4). Samples were collected at 24 hours after treatment. 0.1g of total protein was collected from four-week-old plants. Protein extracted with 150 μL protein extraction buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM EDTA, 0.1% Triton X-100, 0.2% Nonidet P-40, 1 mM PMSF, 1×PIC) and denatured with SDS at 70 ℃ for 10 min. 60 μg protein were separated on 4-12% SDS-PAGE and blotted 1h to nitrocellulose membrane using tank transfer. Blots were blocked with 5% non-fat dry milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 overnight at 4 ℃ with agitation in TBST. 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 RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:2500 in 5% non-fat dry milk in TBST for 1h at RT with agitation. The blot was washed as above and developed for 5 min with SuperSignalTM WEST Pico PLUS Chemiluminescent Substrate from Thermo Scientific. Exposure time was 60 seconds.
Courtesy Msc Min Li, University of South Carolina, USA
||| For other applications, usage on species other than stated above or any other questions, please use the LiveChat option or contact us at firstname.lastname@example.org