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product information
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| background |
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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. |
| immunogen |
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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 |
| antibody format |
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rabbit |
polyclonal |
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affinity purified serum, in PBS pH 7.4 |
lyophilized |
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| quantity |
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180 µg |
for reconstitution add 180 µl of sterile water. |
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| storage |
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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 |
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western blot (WB) |
| related products |
|
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 |
| additional information |
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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
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| recommended dilution |
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1 : 5 000 with standard ECL (WB) |
| expected | apparent MW |
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39 | 45 kDa (Arabidopsis thaliana) |
| confirmed reactivity |
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Arabidopsis thaliana, Capsicum annuum, Lycopersicum esculentum, Pisum sativum, Solanum tuberosum, Zea mays
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| predicted reactivity |
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dicots including Brassica napus, Vitis vinifera, monocots including Oryza sativa, Zea mays |
| not reactive in |
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Chlamydomonas reinhardtii |
| additional information |
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cellular [compartment marker] of mitochondrial matrix |
| selected references |
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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. 
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 SIGMA secondary Anti-Rabbit IgG , Alkaline Phosphatase Conjugate for 1 hour at a dilution of 1:40000 . Blots were developed with the alkaline phosphatase detection system using NBT/BCIP (SIGMA). Courtesy of Bartosz Szabala, Institute of Plant Genetics, Polish Academy of Science.
||| For applications or usage on species others than stated as confirmed above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com
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