PHR1 | Phosphate starvation response 1 (monocots)
AS18 4222 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Hordeum vulgare
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Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
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Samples: purified MBP tag protein, 50 ng, (1), purified MBP-HvTF (MYB-family like PHR1) tag protein 50 ng, (2), purified MBP+HvPHR1, 50 ng, (3). Samples were separated on 13% SDS-PAGE and blotted 1h to PVDF/nitrocellulose using semi-dry transfer. Blot was blocked with 10% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed three times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:50 000 in 10% milk with TBS-T for 1h/RT with agitation. The blot was washed as above and developed for 3 min with chemiluminescent detection reagent. Exposure time was 3 minutes.
Courtesy of Msc. Paweł Sega, Institute of Molecular Biology and Biotechnology, Department of Gene Expression, UAM, Poznań, Poland
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