title: Guaiacol peroxidase zymography for the undergraduate laboratory.
abstract: This laboratory exercise presents a novel way to introduce undergraduate students to the specific detection of enzymatic activity by electrophoresis. First, students prepare a crude peroxidase extract and then analyze the homogenate via electrophoresis. Zymography, that is, a SDS-PAGE method to detect enzyme activity, is used to specifically detect peroxidase activity and furthermore, to analyze the total protein profile. After the assay, students may estimate the apparent molecular mass of the enzyme and discuss its structure. After the 4-h experiment, students gain knowledge concerning biological sample preparation, gel preparation, electrophoresis, and the importance of specific staining procedures for the detection of enzymatic activity.
authors: Center for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Carabobo, Venezuela.
authors: Jeff
authors: J
authors: Wilkesman
authors: Diana
authors: D
authors: Castro
authors: Lellys M
authors: LM
authors: Contreras
authors: Liliana
authors: L
authors: Kurz
mesh_terms: D001671:Biochemistry / Q000193:education / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D008958:Models, Molecular; D008970:Molecular Weight; D009195:Peroxidase / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D015203:Reproducibility of Results; D013334:Students; D013663:Teaching / Q000379:methods*; D014495:Universities
chemical_list: C121048:guaiacol peroxidase; D009195:Peroxidase
keywords: electrophoresis; enzymes; guaiacol peroxidase; teaching protein chemistry; zymography