ICL | Isocitrate lyase
AS09 500 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Ricinus communis
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Purified native ICL from Ricinus communis
64.7 | 65 kDa
Arabidopsis thaliana seeds were germinated according to Benamar et al. (2013). Samples: dry seeds (1), 48h germination (2), 72h germination (3), D4 (4), D7 (5). 20 µg of total protein extracted with SSE buffer containing antiproteases + DTT 10mM + PMSF 1mM and denatured with SB at 90°C for 3 min were separated on 12% Stain Free Gel (Biorad) and blotted 1h to PVDF using tank transfer. Blot was blocked with 3% milk in TBS 0,1% Tween 20 1h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:1 000 for 1h30 at RT. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 15 min in TBS 0,1% Tween20 at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:75 000 in TBST for 45 min. at RT with agitation. The blot was washed as above and developed for 2 min with Clarify Western ECL (Biorad).
Reference: Benamar et al. (2013). Simple system using natural mineral water for high-throughput phenotyping of Arabidopsis thaliana seedlings in liquid culture. Int. J. High Throughput Screen. 4: 1–15.
Courtesy of Dr. David Macherel & Elise Réthoré, IRHS, Angers, France
0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
ICL protein is most abundant in young cotyledons in germinating seedlings, not in rosette leaves or shoots.
Isocitrate lyase (EC=18.104.22.168), localized in glyoxysome is an enzyme involved in mobilization of storage lipis during plant seedling growth. Catalalyzed reaction is a conversion of isocitrate to succinate and glyoxylate. Alternative names of this enzyme: isocitrase, isocitritase, isocitratase, glyoxylate-lyase, isocitrate glyoxylatelyase.
Maeshima et al. (1988). Evidence for no proteolytic processing during transport of isocitrate lyase into glyoxysomes in castor bean endosperm. Plant Cell Physiol. 29:381-384.