The bovine serum albumin (BSA) solution must be diluted to form a series of stan
ID: 823673 • Letter: T
Question
The bovine serum albumin (BSA) solution must be diluted to form a series of standards. The dilutions will be prepared using pipettors and microfuge tips.
? Each sample should have a final volume of 1.0 mL
? Calculate the amount of BSA stock solution (2.0 mg/mL) needed to give a final concentration of BSA shown in the table below.
? Add the correct volume of protein to each tube first, and then add the saline to bring the total volume up to 1.0 mL.
? Once the standards have been diluted, 0.1 mL of each sample will be added to the large glass test tubes for the BCA assay.
TABLE 1: Preparation of BSA Standards Tube #
BSA (2.0 mg/mL)
Water
Final [Protein Stand]
1
0.2 mg/mL
2
0.4 mg/mL
3
0.6 mg/mL
4
0.8 mg/mL
5
1.0 mg/mL
6
1.2 mg/mL
TABLE 1: Preparation of BSA Standards Tube #
BSA (2.0 mg/mL)
Water
Final [Protein Stand]
1
0.2 mg/mL
2
0.4 mg/mL
3
0.6 mg/mL
4
0.8 mg/mL
5
1.0 mg/mL
6
1.2 mg/mL
Explanation / Answer
TABLE 1: Preparation of BSA Standards Tube #
BSA (2.0 mg/mL)
Water
Final [Protein Stand]
1 0.1 ml
0.9 ml 0.2 mg/mL
2 0.2 ml
0.8 ml 0.4 mg/mL
3 0.3 ml
0.7ml 0.6 mg/mL
4 0.4 ml
0.6 ml 0.8 mg/mL
5 0.5 ml
0.5 ml 1.0 mg/mL
6 0.6 ml
0.4 ml 1.2 mg/mL
For case -1 final concn=0.2 mg/ml
N1*V1=N2 *V2
so V1=(N2*V2)/N1=(0.2*1)/2=0.1 ml
so 0.1 ml BSA +0.9 ml saline/water=1.0 ml
similarly calculated for other cases
TABLE 1: Preparation of BSA Standards Tube #
BSA (2.0 mg/mL)
Water
Final [Protein Stand]
1 0.1 ml
0.9 ml 0.2 mg/mL
2 0.2 ml
0.8 ml 0.4 mg/mL
3 0.3 ml
0.7ml 0.6 mg/mL
4 0.4 ml
0.6 ml 0.8 mg/mL
5 0.5 ml
0.5 ml 1.0 mg/mL
6 0.6 ml
0.4 ml 1.2 mg/mL
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