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Featured Individual Experimetns for Human and Animal Physiology

Exp-106 106. Protein Fingerprinting (View Individual Experiment)

A comparison of specific proteins from different species provides a powerful approach for establishing evolutionary relationships and for identifying organisms. A common approach used for this purpose is protein fingerprinting where electrophoretic properties of specific proteins are analyzed in different species. In this exercise, students use the approach to compare the forms of the enzyme lactate dehydrogenase that are found in serum of different mammals. Typical results of this graphic experiment are shown below.

EXP-205 205. Protein Evolution and the Western Blot (View Individual Experiment)

The Western blotting procedure is rapidly replacing conventional methods for identifying and characterizing specific proteins in complex protein mixtures. This technique is used extensively for this purpose in the research laboratory and is increasingly being used in diagnostic medicine for detecting proteins of disease agents such as the structural core proteins of the AIDS virus. Here, students will perform this technique to examine the evolutionary distance between different mammals.

EXP-205P Protein Evolution and the Western Blot (View Individual Experiment)

The Western blotting procedure is rapidly replacing conventional methods for identifying and characterizing specific proteins in complex protein mixtures. This technique is used extensively for this purpose in the research laboratory and is increasingly being used in diagnostic medicine for detecting proteins of disease agents. Here, students will perform this technique to examine the evolutionary distance between different mammals.

EXP-206 206. Affinity Chromatography (View Individual Experiment)

Purified proteins are often needed in the basic research laboratory and for diagnostic and therapeutic procedures. An effective technique for protein purification is affinity chromatography, which exploits a specific interaction between a protein and a complementary binding molecule. In this exercise, students isolate albumin from horse serum by affinity chromatography using a column matrix containing a reactive blue dye which binds specifically to the albumin molecule. They then use electrophoresis to analyze the isolated protein in order to verify the effectiveness of the procedure.

EXP-206P Affinity Chromatography (View Individual Experiment)

Purified proteins are often needed in the basic research laboratory and for diagnostic and therapeutic procedures. An effective technique for protein purification is affinity chromatography, which exploits a specific interaction between a protein and a complementary binding molecule. In this exercise, students isolate albumin from horse serum by affinity chromatography using a column matrix containing a reactive blue dye, which binds specifically to the albumin molecule. They then use electrophoresis to analyze the isolated protein in order to verify the effectiveness of the procedure.

EXP-801 801. Serum Proteins and the Western Press-Blot (View Individual Experiment)

Western blotting is one of the most powerful methods in molecular biology for identifying and characterizing specific proteins in complex protein mixtures. We have now streamlined western-blotting procedures so that the entire analysis can be performed during a single 3-hour, or two 2-hour laboratory sessions. In exercise 801, students use the procedure outlined below to identify albumin, transferrin and gamma globulins in serum and then to study the evolutionary relationships of albumin in vertebrates.

EXP-803 803. Tissue-Specific Isoenzymes in the Cow (View Individual Experiment)

Isoenzymes are different molecular forms of the same enzyme and five major lactate dehydrogenase (LDH) isoenzymes are found in vertebrate tissues. The amounts of the isoenzymes vary in a tissue specific manner and these differences can be readily detected by localizing LDH activity in an agarose gel after electrophoresis of tissue extracts. In this exercise, students prepare a tissue extract from calf thymus and then compare the LDH isoenzyme profile to those from calf serum, heart and muscle.

IND-11P Contractile Proteins from Cow Heart (View Individual Experiment)

In this experiment, students prepare a protein extract from cow heart. They then determine the molecular weights of major contractile proteins by comparing their migration on SDS-polyacrylamide gels to the migration of standard proteins of known size. Students also identify and determine the molecular weight of the major proteins found in milk. This exercise requires 1 three-hour or 2 two-hour lab sessions and a table top centrifuge or microcentrifuge is required. Typical results of the exercise are shown on the gel.

IND-12 Characterization of the Satellite DNA from the Meal Worm (View Individual Experiment)

Satellite DNAs are highly repeated sequences of unknown function. The satellite DNA from the meal worm beetle is remarkable since it represents over 50% of this insects genome. In this exercise, students first isolate DNA from beetle larvae by a simple and safe procedure. They then digest the DNA with EcoR1 and examine the satellite DNA following electrophoresis as shown on the gel pictured below. Suffient materials are provided so that the experiment can be carried out twice by eight groups of students.

IND-26 Localizing Tublin by Immunohistochemistry (View Individual Experiment)

Microtubules are hollow cylinders made up of polymers of the protein tubulin. Microtubules are major components of cilia and flagella, which are tail like projections that are covered by a plasma membrane and extend outwards from the cell. Motile cilia are used for locomotion and food gathering by some protozoa and are found in the lining of the trachea, where their wave like motion propels mucus, dust and other foreign matter out of the lungs.

IND-6 Analysis of a Mutant Hemoglobin Gene (View Individual Experiment)

A mutation is a change in the nucleotide sequence of DNA which leads to an inherited change in an organism. Restriction endonucleases provide valuable tools for characterizing mutations at the DNA level. This principle is illustrated in the exercise where students digest a normal and a mutant gene with EcoR1 and Hae III and then analyze the DNA fragments from each by electrophoresis as shown in the figure below. The gene is from rabbit and codes for the ß-globin chains of hemoglobin.