Goat anti rabbit irdye 800cw 2nd antibody

Detached leaves of mature plants grown under standard conditions for five weeks were floated on water or 2.3 mM lincomycin for 16 h in darkness after which they were illuminated under with high light of 1000 μmol photons m⁻² s⁻¹ for 1.5 h. Thereafter, the water samples were moved to standard growth conditions for recovery. PSII efficiency (F_V/F_M) was measured after a 20 min dark-adaptation using FluorPen FP 110 (Photon Systems Instruments). The amount of D1 protein was measured by western blotting with protein samples collected at indicated timepoints. Total protein samples were isolated by homogenizing frozen leaf material in 100 mM Tris-HCl pH 8.0, 50 mM Na-EDTA, 0.25 mM NaCl, 0.75 (w/v) % SDS, 1 mM DTT followed by 10 min incubation at 68 °C. The extracts were clarified by centrifugation at 12,000 × g for 10 min. Protein concentration was determined using BioRad’s DC Protein assay according to the manufacturer’s protocol. Solubilized protein samples were separated by SDS-PAGE (12% acrylamide, 6 M urea) and immunoblotted with rabbit D1 DE-loop antibody^{91 (link)} used as a 1:8000 dilution. LI-COR Goat anti-rabbit IRDye^® 800CW 2nd antibody was used for detection according to manufacturer’s instructions.