IDH | Isocitrate dehydrogenase

KLH-conjugated peptide 1 and peptide 2 conserved in all higher plants mitochondrial, NAD dependent isocitrate dehydrogenase subunits including Arabidopsis thaliana IDH-I Q8LFC0, At4g35260 and IDH-II P93032, At2g17130

39 | 45 kDa (Arabidopsis thaliana)

Chlamydomonas reinhardtii

Application example

20 µg of total protein from Arabidopsis thaliana leaf extract (1), Arabidopsis thaliana fraction enriched with mitochondria (2), Arabidopsis thaliana pure mitochondria (3), Pisum sativum pure mitochondria (4), Solanum tuberosum pure mitochondria (5),  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, followed by an incubation with a secondary antibody and a series of washes. Blots were developed using chemiluminescent detection reagent.

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

15 µg of total protein stem extract from Lycopersicum esculentum (1), pure mitochondrial fraction isolated from stems of Lycopersicum esculentum (2), pure mitochondrial fraction isolated from stems of Capsicum annuum (3), pure mitochondrial fraction isolated from tubers of Solanum tuberosum (4) were separated on 10% SDS-PAGE and blotted onto nitrocellulose . After blocking with 5% milk in TBST , blots were incubated with the primary antibody at a dilution of 1:1000 in TBST for 1.5h at room temperature. Following incubation and wash steps, blots were incubated with secondary Anti-Rabbit IgG , Alkaline Phosphatase Conjugate for 1 hour at a dilution of 1:40 000. Blots were developed with the alkaline phosphatase detection system using NBT/BCIP.

Courtesy of Bartosz Szabala, Institute of Plant Genetics, Polish Academy of Science, Poland

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).

Cellular [compartment marker] of mitochondrial matrix

AS04 054 | Anti-AOX1/2, marker of inner mitochondrial membrane, rabbit antibodies

AS04 053A | Anti-COXII (plant) marker of inner mitochondrial membrane, rabbit antibodies

AS07 212 | Anti-VDAC1, marker of outer mitochondrial membrane, rabbit antibodies

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.