Purification and Detection of Stumpy Trypanosomes

Stumpy form trypanosomes were purified from blood using DEAE-cellulose DE52 (Whatman) anionic exchange columns [82 (link)] that were preincubated with PSG (44 mM NaCl, 57 mM Na₂HPO₄, 3 mM KH₂PO₄, 55 mM glucose, pH 7.8) warmed to 37°C. Western blotting and antibody concentrations were as described previously [35 (link)]. Anti-EF1α (Millipore) was used at a dilution of 1/7000. Proteins were detected using Enhanced Chemiluminescence reagents (Amersham) and a SRX-101A X-ray developer (Konica Minolta).

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Dewar C.E., MacGregor P., Cooper S., Gould M.K., Matthews K.R., Savill N.J, & Schnaufer A. (2018). Mitochondrial DNA is critical for longevity and metabolism of transmission stage Trypanosoma brucei. PLoS Pathogens, 14(7), e1007195.

Publication 2018

DEAE-cellulose DE52 Anti-EF1α Enhanced Chemiluminescence reagents SRX-101A X-ray developer

Antibody Blood Chemiluminescence Deae cellulose Dilution Glucose Nacl Proteins Trypanosomes X ray

Corresponding Organization : University of Edinburgh

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Variable analysis

independent variables

DEAE-cellulose DE52 (Whatman) anionic exchange columns

dependent variables

Purification of stumpy form trypanosomes from blood

control variables

PSG (44 mM NaCl, 57 mM Na2HPO4, 3 mM KH2PO4, 55 mM glucose, pH 7.8)
Temperature of 37°C

controls

No positive or negative controls were explicitly mentioned in the provided information.

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