TPS1 | Trehalose-6-phosphate synthase 1

Application example

10 µg of total protein from Nicotiana benthamiana leaves expressing AtTPS1-GFP were separated on 10% SDS-PAGE using semi-dry transfer and blotted 1h to PVDF. Crude: non-purified material. Purified: immunoprecipitated with anti-GFP beads. Blots were blocked with 5% skimmed milk powder dissolved in TBS-T (0.1 % Tween 20)  at 4°C ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 for 1h at room temperature (RT) with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 25 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent, according to the manufacturer's instructions. Exposure time was 30 seconds.

Courtesy of Dr. Takeo Sato, Hokkaido University, Japan

TPS1 (Trehalose-6-phosphate synthase 1), EC=2.4.1.15 is an enzyme required for normal embryo development, vegetative growth and transition to flowering. It is involved in the regulation of glucose sensing. Expressed in seedlings, leaves, roots, stems, flowers and siliques. Up-regulated during seed development. Alternative names: alpha-trehalose-phosphate synthase [UDP-forming] 1.