Product Name
ACO1 | Aconitase
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.
Background
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
Reactivity
Arabidopsis thaliana, Solanum lycopersicum
Confirmed reactivity
Arabidopsis thaliana ACO1,ACO2 and ACO3 isoforms, Brassica oleracea, Solanum lycopersicum
Predicted reactivity
Cucurbita maxima, Hordeum vulgare, Nicotiana tabacum, Ricinus communis, Solanum tuberosum, Vitis vinifera, Oryza sativa, Zea mays, Picea sitcHensis, Populus trichocarpa
Not reactive in
Chlamydomonas reinhardii, cyanobacteria
Reconstitution
For reconstitution add 100 µl of sterile water
Expected/apparent MW
98 kDa, Note that ACO1, ACO2 and ACO3 cannot be distinguished in size by standard SDS-PAGE.
Tested applications
Western blot (WB)
Recommended dilution
1 : 5 000 -1 : 10 000 (WB), At higher concentrations the antibody binds aspecifically resulting in non-specific signals around 60 kDa, including Rubisco subunits
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Datasheet 