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Electrophoresis Equipment Not Required
Cross-sections of carrots (large circles), parsnips (small circles), and celery (horseshoe-shaped) were prepared with a razor blade. The proteins at the cut surface of the sections were transferred to nitrocellulose membranes by application of gentle pressure for 10 seconds. The nitrocellulose was then incubated with a color producing peroxidase substrate. Note that the peroxidase is found primarily in the epidermis and in some vascular (phloem) elements.
Locating specific proteins and nucleic acid molecules in tissue sections is an important goal in cell biology. An effective and simple technique for this purpose is tissue printing which permits the localization of specific macromolecules in animal and plant tissues. Here students perform this technique to examine the tissue distribution of the enzyme peroxidase in plants. First, students section carrots, celery, and other vegetables with razor blades and transfer the proteins from the cut sections to nitrocellulose membranes by application of gentle pressure. The enzyme peroxidase is detected on the membranes using a blue color producing peroxidase substrate as shown on the left. All proteins on the membranes are then stained red with a general protein stain which highlights the selective nature of the peroxidase distribution. Students also determine the amount of peroxidase in plant extracts. This 2 hour exercise provides an exciting lesson in plant histology and introduces your student to an important technique used in contemporary cell biology. The exercise was designed for eight groups of students and includes: nitrocellulose membranes, transfer pipets, 200 x extraction buffer, peroxidase standard, chloronapthol, 4M Tris buffer, hydrogen peroxide, and 10 x protein blot stain. Assorted vegetables which can be purchased at your local supermarket and petri dishes are needed, but not provided
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