%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c24%22%3E%3Cbr%20%2F%3E%3C%2Fdiv%3E%0A%3Cp
ID: 826387 • Letter: #
Question
%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c24%22%3E%3Cbr%20%2F%3E%3C%2Fdiv%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3ESample%20%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0ApNPPmM%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%20Volume%0Aof10%20mMPNPP%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%20Volume%20of%0ABuffer%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E0%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.500%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E1%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.025%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%22c1%22%3E2%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A%26nbsp%3B%20%26nbsp%3B%20%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.05%3C%2Fspan%3E%3C%2Fstrong%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E3%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.08%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E4%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.100%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E5%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.150%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E6%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.250%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cstrong%3E%3Cspan%20class%3D%0A%22c1%22%3E7%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%26nbsp%3B%0A.500%3C%2Fspan%3E%3C%2Fstrong%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c1%22%3EDetermine%20the%20amount%20of%20each%0Areactant%3C%2Fdiv%3E%0A%3Cp%3Elisted%20in%20the%20table%20aboveneeded%20for%20a%3C%2Fp%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c2%22%3Etotal%20sample%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c3%22%3Evolume%20of%201.0mL%3C%2Fdiv%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c4%22%3EMATERIALS%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c5%22%3E0.5%20M%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c6%22%3ETris%20Buffer%20pH%208%20with%205%20mM%20MgCl%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c7%22%3Ebovine%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c8%22%3Ealkal%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c9%22%3Eine%20phosphatase%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c10%22%3E(Sigma)%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c11%22%3EM.W.%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c12%22%3E~69%2C000%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c13%22%3EDa%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c14%22%3EStock%20of%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c15%22%3E10mMand%201mM%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c16%22%3Epara-nitrophenyl%20phosphate%20(pNPP)%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c17%22%3Ein%20reaction%20buffer%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c18%22%3E(Sigma)%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c19%22%3EL-phenylalanine%20(10%20mM%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c20%22%3Ein%20reaction%20buffer)%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c21%22%3ESodium%20phosphate%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c22%22%3Emonobasic%3C%2Fdiv%3E%0A%3Cdiv%20dir%3D%22ltr%22%20class%3D%22c23%22%3E(1mM%20in%20reaction%20buffer)%3C%2Fdiv%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0A%3Cp%3E%3Cbr%20%2F%3E%3C%2Fp%3E%0AExplanation / Answer
Experience has shown that enzyme assays used to teach introductory enzyme kinetics should ideally meet a number of criteria. (i) The assay should be reliable and show a prolonged linear rate of reaction to maximize the possibility that inexperienced students will obtain satisfactory data; (ii) given the funding situation at most institutions, the reagents should be inexpensive; and (iii) if preparation time is limited because multiple laboratory classes are offered throughout the week, reagents should be stable. For 2 years we have successfully used variants of one such kinetic assay as the basis for teaching enzyme kinetics in a 2nd-year undergraduate laboratory class. The assay presented in this article involves the hydrolysis of p-nitrophenyl phosphate by a commercially available preparation of the enzyme alkaline phosphatase.
A number of publications describe class experiments with acid phosphatase [1
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