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ICL | Isocitrate lyase

AS09 500 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Ricinus communis

ICL | Isocitrate lyase  in the group Plant/Algal Antibodies / Carbohydrates at Agrisera AB (Antibodies for research) (AS09 500)

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product information
Background

Isocitrate lyase (EC=4.1.3.1), 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.

Immunogen

Purified native ICL from Ricinus communis

Host Rabbit
Clonality Polyclonal
Clone
Purity Ammonium sulfate purified IgG
Format Lyophilized
Quantity 100 µl
Reconstitution For reconstitution add 100 µl of sterile water.
Storage 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.
Tested applications ELISA (ELISA), Western blot (WB)
Related products

collection of antibodies to proteins involved in carbohydrate metabolism

Plant protein extraction buffer

Secondary antibodies

Additional information

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

application information
Recommended dilution 1 : 8000 (ELISA), 1 : 2000 (WB)
Expected | apparent MW

64.7 | 65 kDa

Confirmed reactivity Ricinus communis
Predicted reactivity Arabidopsis thaliana, Cucumis sativus, Gossypium mexicanum, Oryza sativa, Populus balsamifera subsp. trichocarpa
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

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.

Selected references 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.

Application example

Westrn blot using anti-ICL antibodies

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

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