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Understanding Protein Concentration: Dilution Calculations Using Flow Diagrams

This resource provides a clear and simple approach to understanding protein concentration calculations through flow diagrams. It illustrates how to organize your thoughts and apply dilution factors to experiments. The example presented demonstrates the process of calculating the protein concentration of an original sample by working backwards from measured absorbance values. With detailed diagrams showing the dilution process, you will learn how to derive results effectively and accurately, reinforcing concepts in experimental methods.

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Understanding Protein Concentration: Dilution Calculations Using Flow Diagrams

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  1. Flow Diagrams Dr Ron EppingLQB381 Unit Coordinator 1991 - 2012

  2. Simple diagrams help you organise your mind and understand how dilution factors apply to an experiment. Try working backwards using AMOUNTS (g or moles) from the results of the test sample to calculate the AMOUNT in the original sample. Here is an example 100 uL of a protein extract is diluted 1/20 with saline and then assayed for protein as follows: 250 μL of the test sample (diluted extract) is mixed with 100 μL of Reagent A and 500 μL of Reagent B. The solution is then diluted to 3 mL with d.waterand the absorbance measured. From comparison with plotted standards on a standard curve. the 3 mL sample is found to contain 1.5 mg of protein. What is the protein concentration of the original solution in mg/mL?

  3. Flow Diagram: Example Protein Solution PROTEIN SOLUTION 1.0 mL

  4. Flow Diagram: Example 0.1mL Protein Solution PROTEIN SOLUTION 0.1 mL Original Sample 1.0 mL

  5. Flow Diagram: Example 0.1mL Protein Solution PROTEIN SOLUTION 1.9 mL saline 0.1 mL Original Sample 1.0 mL

  6. Flow Diagram: Example 0.1mL 0.25 mL Protein Solution PROTEIN SOLUTION 1.9 mL saline 0.1 mL Original Sample 1.0 mL 2.0 mL

  7. Flow Diagram: Example 0.1mL 0.25 mL Protein Solution PROTEIN SOLUTION 1.9 mL saline 0.25 mL Test Sample 0.1 mL Original Sample 1.0 mL 2.0 mL

  8. Flow Diagram: Example 0.1mL 0.25 mL 0.25 mL Protein Solution PROTEIN SOLUTION 1.9 mL saline 0.25 mL Test Sample 0.1 mL Original Sample 1.0 mL 2.0 mL 0.25 mL

  9. Flow Diagram: Example 0.1mL 0.25 mL 0.25 mL Protein Solution PROTEIN SOLUTION 1.9 mL saline 0.25 mL Test Sample 0.1 mL Original Sample 1.0 mL 2.0 mL 0.25 mL ABSORBANCE ASSAY PROCEDURE 1.5 mg Read from the standard curve + 0.1 mL A + 0.5 mL B + 0.15 mL H2O 1.0 mL

  10. Flow Diagram: Example 0.1mL 0.25 mL 0.25 mL PROTEIN SOLUTION 1.9 mL saline 0.25 mL Test Sample 0.1 mL Original Sample 1.0 mL 2.0 mL 0.25 mL ABSORBANCE ASSAY PROCEDURE 1.5 mg + 0.1 mL A + 0.5 mL B + 0.15 mL H2O CALCULATE USING AMOUNTS: 1.5 x 10 -3 g x 2.0 / 0.25 x 1.0 / 0.1 = 0.12 g in 1 mL original sample = 120 mg/mL 1.0 mL

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