New species reactivity added for anti-PsbO antibodies

Anti-PsbO antibodies, product number: AS06 142-33, detect a PsbO protein in a wide range of different species.

Recently, reactivity on a new species, Eucalyptus grandis, has been experimentally confirmed using the following protocol:

13.5 µg/well of total protein extracted freshly from Eucalyptus sp. leaves. Exact buffer components were: 50 mM HEPES NaOH pH 8.0, 5mM DTT, 15 mM NaHCO3, 20 mM MgCl2, 2 mM EDTA, 4% (v/v) EDTA- free protease inhibitor cocktail (Roche Diagnostics), 4% PVPP (w/v), 20% glycerol (v/v), 1:1 Tris-buffered phenol (v/v) and 100 mM CH3COONa in methanol. Extracts were denatured with SDS beta-Mercaptoethanol sample buffer for 10 min at 90°C. Samples were separated at RT on 12% SDS-PAGE and blotted for 1 h to PVDF, using wet transfer at room temperature (RT). Blot was blocked with 5% non-fat milk for 1 h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 for 1 h at 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 with TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG HRP, Agrisera AS09 602) diluted to 1: 10000 in TBS-T for 1 h at RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent. Exposure time was 12 min.

Anti-PsbO antibodies with reactivity to Eucalyptus

Samples:

1: 13.5 µg of Eucalyptus grandis whole leaf extract clone one (g1)
4: 13.5 µg of Eucalyptus grandis whole leaf extract clone two (g2)
6: 13.5 µg of one clone Eucalyptus grandis x Eucalyptus camaldulensis whole leaf extract (gc)
8: 13.5 µg of one clone Eucalyptus grandis x Eucalyptus tereticornis whole leaf extract (gt)

Presented data courtesy of Ing. Agr. MSc. Matías Nión from Departamento de Biología Vegetal, Facultad de Agronomía, UdelaR. Montevideo, Uruguay.

New species reactivity added for anti-PsbO antibodies

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