Plant/Algal cell biology / Compartment markers / Mitochondria

product information

Plant NADH dependent isocitrate dehydrogenase enzyme is located in mitochondrial matrix. This enzyme is classified as an oxidoreductase and its function is to catalyze a reaction in the citric acid cycle, specifically the sequential dehydrogenation and decarboxylation of isocitrate to form a-ketoglutarate. It removes hydrogens from its substrate, isocitrate. In addition to this process, it functions as a decarboxylase, removing a CO2 from the six-carbon substrate to form a five-carbon product mentioned above as a-ketoglutarate. There are two forms of this enzyme NADP+  and NAD+ dependent.

KLH-conjugated peptide 1 and peptide 2 conserved in all higher plants mitochondrial, NAD dependent isocitrate dehydrogenase subunits including Arabidopsis thaliana IDH-I Q8LFCO and IDH-II P93032

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)

AS04 054 | anti-AOX1/2 rabbit antibody, marker of inner mitochondrial membrane

AS04 053A | anti-COXII rabbit antibody marker of inner mitochondrial membrane

AS07 212 | anti-VDAC1 rabbit antibody marker of outer mitochondrial membrane

Peptide used to elicit this antibody is not conserved in NADPH dependent anzymes, partially conserved across eukaryotic Idh subunits. Some conservation across bacterial which contain the NAD-dependent form of Idh (as opposed to the NADP-dependent form).

application information

1 : 5 000 with standard ECL (WB)

39 | 45 kDa (Arabidopsis thaliana)

Arabidopsis thaliana, Pisum sativum, Solanum tuberosum, Zea mays

dicots including Brassica napus, Vitis vinifera, monocots including Oryza sativa, Zea mays

Chlamydomonas reinhardtii

cellular [compartment marker] of mitochondrial matrix

to be added when available

application example

20 µg of total protein from (1) Arabidopsis thaliana leaf extract, (2) Arabidopsis thaliana  fraction enriched with mitochondria, (3) Arabidopsis thaliana  pure mitochondria, (4) Pisum sativum pure mitochondria, (5) Solanum tuberosum pure mitochondria were separated on  4-12% SDS-PAGE and blotted to nitrocellulose. Blots were blocked immediately following transfer in 5% milk powder in TBS. Blots were incubated in the primary antibody at a dilution of 1: 5 000 for 1h at room temperature with agitation. Blots were developed using ECL reagent (GE Healthcare).

* Band detected at ca. 90 kDa is suspected to be a dimmer of Idh, since this band is depleted upon peptide competition experiment.