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ICL | Isocitrate lyase

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

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

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Product name, number (Agrisera, Sweden)

Datasheet Product citations Protocols Add review

Product Information

Immunogen

Purified native ICL from Ricinus communis

Host Rabbit
Clonality Polyclonal
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)
Recommended dilution 1 : 8000 (ELISA), 1 : 2000 (WB)
Expected | apparent MW

64.7 | 65 kDa

Reactivity

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

Application examples

Application examples 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

Additional information

Additional information

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.

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Background

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.

Product citations

Selected references Réthoré et al. (2019). Arabidopsis seedlings display a remarkable resilience under severe mineral starvation using their metabolic plasticity to remain self-sufficient for weeks. Plant J. 2019 Mar 22. doi: 10.1111/tpj.14325.
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.

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