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a.) if we add 100 microliters of 0.5 mg/ml Con A and 100 microliters of 1 M mann

ID: 9152 • Letter: A

Question

a.) if we add 100 microliters of 0.5 mg/ml Con A and 100 microliters of 1 M mannose to 800 microliters of a dilute suspension of red blood cells, what should we expect to happen? Explain why we see these results. (Describe what is happening at the cellular and/or molecular level rather than what you actually see with your naked eye).

b.) if you have a Con A sample with an A280 reading of 1.513 at a 1:5 dilution, what is the mg/ml of your Con A sample?

c.) how much of this original sample will you need to set up a 5 mg/ml sample in 1X Con A buffer at a final volume of 500 microliters?

d.) how much of this original sample will you need to set up a 2 mg/ml samle in 1X Con A bugger at a final volume of 300 microliters?

Explanation / Answer

b.) The molar absorption coefficient of jack bean con A = 32430. The mol.mass = 25572 g --> 25572 g/L = 32430 Abs. units 1 g/L = 1 mg/mL = 32430/25572 = 1,268 A.units. An absorption of 1,513 = 1*1,513/1,268 mg/mL = 1,2 mg/mL And as it was diluted by 1:5, the original concentration is 6 mg/mL