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4. Read the absorbance at 595nm of the blank and test solutions. Calculate the protein concentration (in mg/ml) of an unknown sample with an absorbancy value of 0.259 using the standard curve below. 1.5 1.4 1.3 1.2 1.1 1.0 E 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 1.6 BSA concentration (mg/ml) Absorbancy 595nm

4. Read the absorbance at 595nm of the blank and test solutions. Calculate the protein concentration (in mg/ml) of an unknown sample with an absorbancy value of 0.259 using the standard curve below. 1.5 1.4 1.3 1.2 1.1 1.0 E 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 1.6 BSA concentration (mg/ml) Absorbancy 595nm

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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QUESTION 3 An unknown protein was determined using the following method: Bradford method Reagents Stock Bradford reagent: Dissolve 100 mg Coomassie Brilliant Blue G250 in a mixture consisting of 100ml of 85% phosphoric acid, 50ml of 95% ethanol and 50ml 1M NaOH. Store at 4°C until precipitation occurs, at which point it is discarded. Working Bradford reagent: Prepare fresh by diluting 10ml of stock Bradford reagent to 250ml with distilled water. Stock bovine serum albumin (BSA) solution (10mg/ml): Dissolve 0.2g BSA in 20ml distilled water. Working BSA concentration range: 0.5 – 1.25 mg/ml Method 1. Prepare the following test tubes: Table 1: Preparation of test tubes for the Bradford method Blank Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 Unknown Distilled H20 1.0 ml 0.9ml 0.9ml 0.9ml 0.9ml 0.9ml 0.8ml 0.25 mg/ml BSA 0.1 ml 0.5 mg/ml BSA 0.1 ml 0.75 mg/ml BSA 0.1 ml 1.0 mg/ml BSA 0.1 ml 1.25 mg/ml BSA 0.1ml Unknown 0.2ml protein Working 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml Bradford 2. After each addition of the working Bradford reagent, thoroughly mix the resultant solution. 3. Incubate the solution at room temperature for 5 min.
Transcribed Image Text:QUESTION 3 An unknown protein was determined using the following method: Bradford method Reagents Stock Bradford reagent: Dissolve 100 mg Coomassie Brilliant Blue G250 in a mixture consisting of 100ml of 85% phosphoric acid, 50ml of 95% ethanol and 50ml 1M NaOH. Store at 4°C until precipitation occurs, at which point it is discarded. Working Bradford reagent: Prepare fresh by diluting 10ml of stock Bradford reagent to 250ml with distilled water. Stock bovine serum albumin (BSA) solution (10mg/ml): Dissolve 0.2g BSA in 20ml distilled water. Working BSA concentration range: 0.5 – 1.25 mg/ml Method 1. Prepare the following test tubes: Table 1: Preparation of test tubes for the Bradford method Blank Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 Unknown Distilled H20 1.0 ml 0.9ml 0.9ml 0.9ml 0.9ml 0.9ml 0.8ml 0.25 mg/ml BSA 0.1 ml 0.5 mg/ml BSA 0.1 ml 0.75 mg/ml BSA 0.1 ml 1.0 mg/ml BSA 0.1 ml 1.25 mg/ml BSA 0.1ml Unknown 0.2ml protein Working 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml 4.0ml Bradford 2. After each addition of the working Bradford reagent, thoroughly mix the resultant solution. 3. Incubate the solution at room temperature for 5 min.
3. Incubate the solution at room temperature for 5 min. 4. Read the absorbance at 595nm of the blank and test solutions. Calculate the protein concentration (in mg/ml) of an unknown sample with an absorbancy value of 0.259 using the standard curve below. 1.5 1.4 1.3 1.2 1.1 1.0 09 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 1.6 BSA concentration (mg/ml) Absorbancy 595nm
Transcribed Image Text:3. Incubate the solution at room temperature for 5 min. 4. Read the absorbance at 595nm of the blank and test solutions. Calculate the protein concentration (in mg/ml) of an unknown sample with an absorbancy value of 0.259 using the standard curve below. 1.5 1.4 1.3 1.2 1.1 1.0 09 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 1.6 BSA concentration (mg/ml) Absorbancy 595nm
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