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Lab 3. PROTEIN DETERMINATION

INTRODUCTION

Reading in Biology, 6th ed. By Campbell: “PROTEINS-MANY STRUCTURES, MANY FUNCTIONS” p. 71-80 “Blood is a connective tissue with cells suspended in plasma” p. 882-884. Protein is a major and indispensable class of cellular macromolecules. Accordingly, measuring protein concentrations in an amazingly diverse array of experimental contexts (How many grams of protein are there in a 27 gram serving of Fruit Loops?) continually engages the attention, and taxes the resources, of many investigators. The two essential and frequently cited basic references on methods for general protein determination are: Layne, (1957) and Peterson, (1983). These articles are on hard copy reserve for the class at the Science Library in the folder for this exercise. Two things impressed me as I read these papers. First, the methods described are all optical methods. That is, measurement of protein concentration relies on optical properties (absorption or turbidity) of protein solutions and these optical properties are measured with a device called a spectrophotometer (or colorimeter). Other types of methods (gravimetric, volumetric, etc.) for protein determination exist, such as the classic chemical method of Kjeldahl, but other such methods were not even discussed in these papers. Optical methods of biochemical analysis are convenient, sensitive and versatile. We will use optical methods for several additional exercises later in this course. Second, these papers described no fewer than 11 different methods for protein determination, even though they limited themselves to a presentation of optical methods rather than chemical methods. The existence of so many different methods must mean, I'm very, very sorry to say, that no single method is appropriate for all experimental needs in all laboratory contexts. Thus, before tackling the job of conducting protein assays an investigator is faced with the rather more challenging prospect of choosing the...

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