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15 14 13 12 11 1.0 0.9 0.8 0.7 0.6 05 0.4 0.3 02 0.1 00 0.0 0.1 0.2 03 0.4 0.5 0.6 0.7 08 09 10 11 12 13 14 15 16 BSA concentration (mg/ml) Absorbancy 595nm

15 14 13 12 11 1.0 0.9 0.8 0.7 0.6 05 0.4 0.3 02 0.1 00 0.0 0.1 0.2 03 0.4 0.5 0.6 0.7 08 09 10 11 12 13 14 15 16 BSA concentration (mg/ml) Absorbancy 595nm

Curren'S Math For Meds: Dosages & Sol
Curren'S Math For Meds: Dosages & Sol
11th Edition
ISBN:9781305143531
Author:CURREN
Publisher:CURREN
Chapter10: Reconstitution Of Powdered Drugs
Section: Chapter Questions
Problem 7.6P
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1.5 1.4 1.3 1.2 1.1 1.0 0.9 0.8 0.7 0.6 05 0.4 0. 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:1.5 1.4 1.3 1.2 1.1 1.0 0.9 0.8 0.7 0.6 05 0.4 0. 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
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 folloving 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 0.9ml 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 0.1ml 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. 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 0.842 using the standard curve below.
Transcribed Image Text: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 folloving 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 0.9ml 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 0.1ml 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. 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 0.842 using the standard curve below.
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