Detection of tubulin-binding proteins by an overlay assay.
A blot overlay technique to detect tubulin-binding proteins has been developed. It involves the fractionation of the putative tubulin-binding proteins by gel electrophoresis, their transfer to nitrocellulose paper, and the incubation of the nitrocellulose sheet with purified tubulin. The proteins which bind tubulin are recognized by monoclonal anti-tubulin antibodies. Proteins which associate with microtubules through a direct interaction with tubulin are easily detected by this procedure, which has been tested for both known microtubule-associated proteins and newly reported tubulin-binding proteins.
Kremer
L
L
Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma, Madrid, Spain.
Domínguez
J E
JE
Avila
J
J
D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification*; D015151:Immunoblotting / Q000379:methods*; D008869:Microtubule-Associated Proteins / Q000302:isolation & purification; D014404:Tubulin / Q000378:metabolism*
D002352:Carrier Proteins; D008869:Microtubule-Associated Proteins; D014404:Tubulin