SPS | Sucrose phosphate synthase, global
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KLH-conjugated synthetic peptide derived from conserved region within plant SPS protein sequences, including Arabidopsis thaliana isoforms 1F Q94BT0, 2F, 3F and 4F. Oryza sativa Q67WN8, Solanum tuberosum Q43845
120 | 120-130 kDa (fragments of 30-90 kDa may be detected)
10 µg of total leaf protein from (1) A.thaliana,(3) Zea mays and (4) Hordeum vulgare extracted with PEB (AS08 300) as well as 10 µg cytosolic protein from (2) A.thaliana were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1.5h (30V) to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-SPS (AS03 035A, 1:2000, 1h) and secondary anti-rabbit (1:20 000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. Antibody incubations where followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent detection reagent, using a Fuji LAS-3000 CCD (90s, high sensitivity).
Peptide used to elicit anti-SPS antibodies is perfectly conserved in all isoforms of SPS in plants.
SPS (sucrose phosphate synthase, EC 22.214.171.124) is the key enzyme of carbon flux into sucrose fixation in plants. It catalyzes the synthesis of sucrose-phosphate from UDP-glucose and fructose-6-phosphate predominantly in the cytosol of sucrose-source leaf tissue.
Zhang et al. (2014). Heterologous expression of AtPAP2 in transgenic potato influences carbon metabolism and tuber development. FEBS Lett. 2014 Aug 27. pii: S0014-5793(14)00621-8. doi: 10.1016/j.febslet.2014.08.019.