6. Calculate the diluted concentration (in mg/mL) of standard protein in each tube S1 – S5 using the concentration of the standard protein solution and the volumes used to prepare the diluted protein standards. Record these concentrations on the report sheet. Protein Standards Unknowns Tube Number Blank S1 S2 S3 S4 S5 U1 U2 Volume of Standard Protein 0.00 mL 0.10 mL (100 ????L) 0.20 mL (200 ????L) 0.40 mL (400 ????L) 0.60 mL (600 ????L) 0.80 mL (800 ????L) 1.00 mL (1000 ????L) Unknown In Each Tube Volume of 1% NaCl 1.00 mL (1000 ????L) 0.90 mL (900 ????L) 0.80 mL (800 ????L) 0.60 mL (600 ????L) 0.40 mL (400 ????L) 0.20 mL (200 ????L) Volume of Biuret Reagent 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL
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First, we need to find the total volume of each tube (S1-S5). To do this, we add the volume of the standard protein and the volume of 1% NaCl. Show more…
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Calculate the diluted concentration (in mg/mL) of standard protein in each tube S1 – S5 using the concentration of the standard protein solution and the volumes used to prepare the diluted protein standards. Record these concentrations on the report sheet. Protein Standards Unknowns Tube Number Blank S1 S2 S3 S4 S5 U1 U2 Volume of Standard Protein 0.00 mL 0.10 mL (100 ̄̄̄̄L) 0.20 mL (200 ̄̄̄̄L) 0.40 mL (400 ̄̄̄̄L) 0.60 mL (600 ̄̄̄̄L) 0.80 mL (800 ̄̄̄̄L) 1.00 mL (1000 ̄̄̄̄L) Unknown In Each Tube Volume of 1% NaCl 1.00 mL (1000 ̄̄̄̄L) 0.90 mL (900 ̄̄̄̄L) 0.80 mL (800 ̄̄̄̄L) 0.60 mL (600 ̄̄̄̄L) 0.40 mL (400 ̄̄̄̄L) 0.20 mL (200 ̄̄̄̄L) Volume of Biuret Reagent 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL 4.00 mL
Jenny W.
Determination of protein concentration You want to determine the protein concentration in an unknown sample by use of the biuret method. The standard solutions needed were prepared by mixing the bovine serum albumin (BSA) stock solution, water, and the biuret reagent, as can be seen in the data table below. (The concentration of the BSA stock solution was 8.00 mg/mL). The solutions were left at room temperature for 30 minutes for the complete color development. The absorbance (at 540 nm) of each of the standard solutions was determined with a spectrophotometer using the first solution (with 0.000 mg/mL protein) as the blank (control), as shown below. test tube # volume of BSA volume of water volume of biuret reagent final protein conc. Abs540 1 0.00 mL 2.00 mL 2.00 mL 0.000 mg/mL 0.00 2 0.20 mL 1.80 mL 2.00 mL 0.075 3 0.40 mL 1.60 mL 2.00 mL 0.162 4 0.60 mL 1.40 mL 2.00 mL 0.230 5 0.80 mL 1.20 mL 2.00 mL 0.333 6 1.00 mL 1.00 mL 2.00 mL 0.403 2.00 mL of the unknown sample in a cuvette was mixed with 2.00 mL of the biuret reagent. After 30 minutes, the absorbance of the unknown sample in the cuvette was measured to be 0.275. Your lab report should include the following: (1) Complete the data table shown above. (The calculations of the final protein concentrations should be shown below the table.)
Tube1 info - Standard protein solution: 80uL - Water: 120uL - Total volume in tube: 200uL - The standard protein was used at a concentration of 500ug/mL. I got an answer for the concentration of the standard: Concentration (500ug/mL) × Volume (80uL) = Concentration (final) × Volume (final) (200ug/mL) --> 500ug/mL × 0.08mL = Concentration (final) × 0.2mL --> 40ug = Concentration (final) × 0.2mL --> 40ug / 0.2mL = Concentration (final) --> Concentration (final) = 200ug/mL Is this question different from my concentration of the standard answer?
Rebecca C.
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