Molluscan shell matrix characterization by preparative SDS-PAGE.
The glycoproteinaceous constituents of molluscan shell matrices usually resist chromatographical fractionation. We describe a protocol that overcomes this difficulty and permits collection of a large amount of shell proteins for further in vitro characterization. After dissolution of the mineral phase, the glycoproteins are fractionated "blind" on a preparative electrophoresis. They are subsequently detected with a polyclonal antibody raised against the whole matrix.
Marin
Frédéric
F
IsoTis, NV., ProfBronkhorstlaan, 10, Gebouw D, 3723 MB Bilthoven, The Netherlands.
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D005591:Chemical Fractionation / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005620:Freund's Adjuvant / Q000276:immunology; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D007114:Immunization / Q000379:methods; D008974:Mollusca / Q000737:chemistry*; D011489:Protein Denaturation; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*
D000906:Antibodies; D006023:Glycoproteins; D012967:Sodium Dodecyl Sulfate; D005620:Freund's Adjuvant