Plant/Algal cell biology / Physcomitrella patens

product information

GDP-L-Galactose Phosphorylase is a histidine triad (HIT) enzyme of the Smirnoff-Wheeler pathway of ascorbic acid synthesis in plants. Encoded by VTC2 gene. The enzyme catalyzes the conversion of GDP-L-galactose to L-galactose 1-phosphate in a reaction that consumes inorganic phosphate and produces GDP.

KLH-conjuated synthetic peptide derived from known GDP-L-Galactose Phosphorylase sequences, including Arabidopsis thaliana Q8LKQ7 and Chlamydomonas reinhardtii 

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

western blot (WB)

to be added when available

application information

1 : 1200 with alkaline phosphatase, 1: 3000 with regular ECL (WB)

51 | 50  kDa

Arabidopsis thaliana, Chlamydomonas reinhardtii

dictos: Nicotiana tabacum, Sorghum bicolor, monocots: Zea mays, Oryza sativa, moss: Physcomitrella patens

no confirmed exceptions from predicted reactivity known in the moment

to be added when available

to be added when available, antibody available in July 2010

application example

10 ug of total soluble protein from Chlamydomonas reinhardtii  was  separated on 7.5% SDS-PAGE and blotted 1.5h to nitrocellulose at 1.5 mA/cm2 constant current. Blots were blocked immediately following transfer in PBS-0.3% Tween 20 + 1% dried milk overnight at room temperature (RT) with agitation. Blots were incubated in the primary antibody AS10 723 at a dilution of 1: 1200 for 4 h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly once with PBS-T+1%milk, followed by washing 3 times for 5 min in the same at RT with agitation. Blots were incubated in secondary antibodies (goat anti-rabbit IgG AP conjugated) diluted in PBS-T+1%milk to 1:3000 for 1 h at RT with agitation. The blots were washed 2 x 5 mins with PBS-T+1% milk as above, then rinsed with TBS and color developed (approx 5 minutes).

Courtesy Dr. Dudley Page, UCLA