How to Measure Antibody Concentration
In medicine, and in scientific experiments, it is often necessary to experimentally determine the concentration of an antibody protein in a solution. Science suppliers, for most commonly studied antibodies, provide kits through which the concentration of an antibody can be measured by adding a reactant to a solution and measuring the absorbance of that solution in a spectrophotometer. The absorbance will be proportional to the concentration of antibody.
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Instructions
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Purchase a kit, from a science supplier online, designed to allow you to measure the concentration of an antibody in a solution.
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Add the reagents in the kit to your solution of antibody, in the proportions instructed by the kit. For instance, you might be instructed to add 100 ml of antigen to 1 ml of the antibody solution --- in other words, add 100 ml of a protein that will bind specifically to the antibody --- along with 100 ml of a fluorescent dye that will bind to the antigen. Use the proportions recommended by the manufacturer and to add reagents with precision.
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Pipette 100 ml of your solution into a spectrophotometer plate, and place your plate into the spectrophotometer. Read the contents of your plate, using the spectrophotometer, to obtain an absorbance value for your solution.
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Compare your absorbance value with the standard curve, provided in your kit, of absorbance versus concentration. Plug your absorbance value into the equation describing your standard curve, to calculate an approximation of the antibody concentration of your solution.
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