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Available ordering format
Lyophilized
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Immunogen
Arabidopsis ACO1 (AT4G35830, Q42560), codon 120 - 898 (C-terminus), was cloned in fusion with a N-terminal 6xHis tag, and over-expressed in E. coli. All recombinant protein accumulated in inclusion bodies, which were purified by centrifugation and solubilised in 6 M guanidine-HCl. The protein was refolded by dilution in 100 mM Tris-HCl 8.5, 10% (v/v) glycerol, 2 mM dithiothreitol, and concentrated prior to immunisation.
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Raised in
Rabbit
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Clonality
Polyclonal
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Clone
Polyclonal
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Purification
Serum
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How to reconstitute
For reconstitution add 100 µl of sterile water.
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Storage condition
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.
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Verified applications
western blot (WB)
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Connected products
collection of antibodies to proteins located in mitochondriaPlant protein extraction bufferSecondary antibodies
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Recommended dilutions for use
1:5 000 - 1:10 000 with standard ECL (WB). At higher concentrations the antibody binds aspecifically resulting in non-specific signals ≤ 60 kDa, including Rubisco subunits.
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Molecular weight expected аpparent
98 kDa. Note that ACO1, ACO2 and ACO3 cannot be distinguished in size by standard SDS-PAGE.
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Verified reactivity
Arabidopsis thaliana ACO1,ACO2 and ACO3 isoforms, Solanum lycopersicum
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Possible reactivity
dicots including:Cucurbita maxima, Nicotiana tabacum, Ricinus communis, Solanum tuberosum, Vitis vinifera,monocots: Oryza sativa, Zea mays, trees: Picea sitchensis, Populus trichocarpa
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No reactivity
Chlamydomonas reinhardii
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Supplementary information
The antibody recognises all three Arabidopsis aconitase isoforms (ACO1, ACO2 and ACO3, see Bernard et al 2009). Possible differences in affinity have not been precisely quantified. Sensitivity threshold is between 2 and 10 ng for WB / ECL (see figure). Antibodies will recognize aconitase isoforms in denaturing and native gel electrophoresis.
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References
Rurek et al. (2015). Biogenesis of mitochondria in cauliflower (Brassica oleracea var. botrytis) curds subjected to temperature stress and recovery involves regulation of the complexome, respiratory chain activity, organellar translation and ultrastructure. Biochim Biophys Acta. 2015 Jan 21. pii: S0005-2728(15)00016-X. doi: 10.1016/j.bbabio.2015.01.005. Setién et al. (2014). Root phosphoenolpyruvate carboxylase and NAD-malic enzymes activity increase the ammonium-assimilating capacity in tomato. J Plant Physiol. 171:49-63.Birke et al. (2012). Cysteine biosynthesis, in concert with a novel mechanism, contributes to sulfide detoxification in mitochondria of Arabidopsis thaliana. Biochem J. May 2, ahead of print.Bernard et al. (2009). An allelic mutant series of ATM3 reveals its key role in the biogenesis of cytosolic iron-sulfur proteins in Arabidopsis. Plant Physiol. 151:590-602.
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Scientific context
Aconitase is a single subunit enzyme of the tricarboxylic acid cycle (or Krebs cycle) in the mitochondria. A cytosolic isoform is also part of the glyoxylate cycle. Aconitase catalyzes the dehydration / hydration of citrate to iso-citrate, via cis-aconitate as an intermediate. The reaction is facilitated by an iron-sulphur cluster in the active site of the enzyme. The iron-sulphur cluster is somewhat unstable, especially under oxidative stress, and loss of the cofactor leads to degradation of the protein.Alternative names: ACO, citrate hydro-lyase 1,2,3
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Notes
Arabidopsis expresses three highly similar aconitase isozymes (ACO1/ AT4G35830, ACO2/AT4G26970 and ACO3/AT2G05710), of which ACO1 is the cytosolic isoform, while ACO2 and ACO3 are predominantly located in the mitochondria (Arnaud et al 2007, Bernard et al 2009). The combined abundance and activity of the mitochondrial aconitases is about 3 times higher than the cytosolic pool (Bernard et al 2009).The Arabidopsis isoforms are more similar in amino acid sequence to mammalian iron-regulatory proteins (IRP-1) than to the mammalian and yeast mitochondrial aconitases.
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Protein number
Q42560
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TAIR number
AT4G35830
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