Question

Do you mind checking my work please? I feel like there's something wrong with question 2 and 3 but I can't quite put my finger on it. If it is wrong, could you please explain it in detail? Also, need help filling out the table for the mass of the enzyme. Thank you for your help!

1. Math/Calculation: What is the concentration of substrate in the reaction cocktail? C 50 mM pNPP 50mM -0.05 moVL pNPP Vi-500 μ1-0.0005 L Reaction Cocktail V2-4000 μ-0.004 L Reaction Cocktail (0.05 mo/L)(0.0005 L) C2(0.004L) 2.5 x 10-5 mol pNPP C20.004L) 6.25 x 103 M 6.25 mM pNPP in 4000 μ1 Reaction Cocktail 2. Math/Calculation: What is the concentration of substrate in the reaction tube? [Hint: each Tube 1-6 had the same concentration of substrate, so this only needs to be solved once.] C1-6.25 mM pNP 6.25 mM 0.00625 moVL pNPP Vi-4000 μ1 reaction cocktail = 0.004 L V2-400 μ1 per tube-0.0004 L (0.00625 molVL)(0.004 L)- C(0.0004L) 2.5 x 10 5 mol-Ca(0.00041.) 0.0625 M-62.5 mM of pNPP in 400 μ1 Reaction Cocktail (per tube 1-6) 3. Math/Calculation: How many moles of substrate are in the reaction tube? M-mo/L 62.5 mM pNPP 0.0625 mol/L pNPP 400 μ1 = 0.0004 L Reaction Cocktail (per tube 1-6) (0.0625 mol/L pNPP)(0.0004 L) -2.5 x 10-5 mol pNPP Table 1: Reaction Cocktail Solution/ChemicalVolume to add 1 M acetate buffer pH 5.7 0.1 M MgCl2 500 μι 500ul 50 mM pNPP S Wostrate 500u 2500ul Water Total volume 4000 | 11. Record the concentration of the enzyme from the instructor. o.2 mgIm Table 2: Acid Phosphatase Assay Protocol # | Cocktail ul Tube | Reaction | 0.10%BSA | Enzyme (ul) | Mass enzyme | Time | A410 Wait (mg) | | 100 μ1 80μ1 60 ul 40 I 20 μι O ul 15 min Smin.074 0.00 400 ul 400 μ! 40011 20 ul 40 ul ー60111 ismin 10.11 o.2IS 4 5 6 400 μ! 400 μι 400 μ! 80 ul 100 ul ismin | Ο.SSS

Answer 1

Woking fom 01 02 03 04 05 hagle 06 1000 Notes:

2, X16 3 Similby 0 20 40 t-X10- go 100 3