One step flow-through adsorptive purification of tubulin from tissue homogenate.
Tubulin, a potential target for anti-cancer drugs, has been purified in one step and obtained as flow-through fraction directly from an extract of a mammalian brain tissue by adsorption chromatography on H-CELBEADS, an indigenously developed rigid, superporous cross-linked cellulose based weakly hydrophobic adsorbent. The fibrous polymerized tubulin mass passed through the H-CELBEADS bed while the associated proteins were separated by adsorption. The final tubulin preparation was obtained free from other proteins as seen on SDS-PAGE. Purified tubulin was obtained in a yield of about 29 mg/100 g brain, and its bioactivity, evaluated through its ability to bind colchicine, was found to be preserved.
Anil
Annamma
A
Chemical Engineering Division, Mumbai University Institute of Chemical Technology, Matunga, Mumbai 400019, India.
Pandit
Reena
R
Indap
Madhavi
M
Lali
Arvind
A
D000327:Adsorption; D000818:Animals; D001923:Brain Chemistry; D002482:Cellulose; D002845:Chromatography / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006041:Goats; D057927:Hydrophobic and Hydrophilic Interactions; D009391:Nephelometry and Turbidimetry; D020836:Protein Structure, Quaternary; D013696:Temperature; D014404:Tubulin / Q000302:isolation & purification*
D014404:Tubulin; D002482:Cellulose