Anti psba

Protein extracts from plants were subjected to SDS-PAGE followed by either Coomassie Brilliant Blue (CBB) staining or immunoblot analysis. Immunoreactive signals were detected using an ECL detection system (GE Healthcare) with the following antibodies: anti-RBCL (1:1000 dilution), anti-RBCS (1:1000), anti-PSBA (1:5000), and anti-PGL35 (1:1000) (Agrisera, Sweden).

For immunoblotting, proteins were separated by 12% SDS-PAGE and
electrotransferred to Immobilon-P PVDF membrane (Millipore). The membranes were
blocked for 2 h at room temperature with 5% BSA in PBS-Tween buffer (137 mM NaCl,
2.7 mM KCl, 8.1 mM NaH₂PO₄, 1.5 mM
KH₂PO₄ and 0.1% Tween-20).
Immunoblots were conducted overnight at 4°C. Antibodies used in this study
included anti-Histone H3, anti-UDPase, anti-PsbA (AgriSera AB [84 ]), and anti-RIN. After washed with PBS-Tween
(3 × 10 min), the membranes were treated with corresponding secondary antibodies
conjugated to horseradish peroxidase. The immunoreactive bands were visualized by
a chemiluminescence detection kit (SuperSignal®, Pierce Biotechnology
[85 ]).

Total proteins extraction was performed as previously described^{50 (link)} isolated from wild type and wfsl1 at seedling and tillering stage. The tissues were ground in liquid nitrogen and thawed in extraction buffer [50 mM Tris–HCl pH 7.5, 150 mM NaCl, 10% glycerol (v/v), 0.1% Nonidet P-40, 1 mM DTT, 1 mM PMSF, and 1x complete protease inhibitor cocktail (Roche)] for 15 min on ice. The supernatant was collected by centrifugation at 12,000 g for 10 min at 4 °C. Total proteins were separated by SDS-PAGE gels (8%), transferred to the polyvinylidene difluoride (PVDF) membranes (GE Healthcare), blotted with different primary antibodies, detected with ECL prime (GE Healthcare). Anti-WFSL1 antibody was obtained from Shanghai Youke Biotechnology (http://www.youke-ab.cn/) and other antibodies such as anti-PsaA (LOC_Osp1g00340.1, Cat:AbP80033-A-SE), anti-AtpB (LOC_Os10g21266.1,Cat:AbP80331-A-SE), anti-RCA (LOC_Os11g47970.1, Cat:AbP80246-A-SE) and anti-RpoA (LOC_Osp1g00660.1, Cat:AbP80103-A-SE) from Beijing Protein Innovation (http://www.proteomics.org.cn/) and anti-PsbA (LOC_Osp1g00110.1, Cat:AS05084) from Agrisera.