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Quantitation and characterization of the microtubule associated MAP2 in porcine tissues and its isolation from porcine (PK15) and human (HeLa) cell lines |
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| Authors: | Manuel M Valdivia Jesús Avila Javier Coll Camilo Colaço Ignacio V Sandoval |
| Institution: | 1. Centro de Biología Molecular (CSIC). Universidad Autónoma. Canto Blanco. Madrid-34. Spain.;2. Servicio de Inmunología. Centro S. Ramón y Cajal. Madrid-34. Spain.;3. Division of Biology. California Institute of Technology. Pasadena, California 91125. USA. |
| Abstract: | A radioimmunoassay developed for the microtubule associated protein MAP2 shows that this protein, or related polypeptides are present in all the porcine tissues studied. Nervous tissues (brain, 11 μg MAP2/mg protein; cerebellum, 9.7 μg MAP2/mg protein) contain much higher levels of MAP2 than non-nervous tissues (kidney, 104 ng MAP2/mg protein; lung 89 ng MAP2/mg protein; spleen 66 ng MAP2/mg protein; thyroid 21 ng MAP2/mg protein; liver 9.7 ng MAP2/mg protein). A heat resistant protein doublet of 300,000 with the ability to promote microtubule polymerization has been purified from pig kidney cells by affinity chromatography using MAP2 antibodies. Using a similar purification method a protein of 200,000 daltons has been isolated from Hela cells. |
| Keywords: | SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis EGTA ethyleneglycol-bis-(β-amino ethylether)N N′-tetra-acetic acid MES 2-(N-morpholino)ethanesulfonic acid Tris Tris(hydroxymethyl) amino methane EBSS Eagles basic salt solution |
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