References spectrophotometry 1 segel ih 1976

This preview shows page 8 - 10 out of 28 pages.

We have textbook solutions for you!
The document you are viewing contains questions related to this textbook.
Understanding Normal and Clinical Nutrition
The document you are viewing contains questions related to this textbook.
Chapter 22 / Exercise 2
Understanding Normal and Clinical Nutrition
Rolfes/Whitney
Expert Verified
References Spectrophotometry 1. Segel, I.H. (1976) Biochemical Calculations, 2 nd ed., Chapter 5, John Wiley & Sons, New York. 2. Freifelder, D. (1982) Physical Biochemistry , 2nd ed., Chapter 14, pages 494-500, 504-507, 511-512. W. H. Freeman and Co., San Francisco. 3. Cooper, T. (1977) The Tools of Biochemistry , Chapter 2, John Wiley & Sons, New York. Spectra and Molar Absorptivities 1. Long, C. ed. (1961) Biochemist's Handbook , pp. 81-82, D. Van Nostrand Co., Inc., New York. 2. Kirschenbaum, D. M. (1972) Atlas of Protein Spectra in the Ultraviolet Visible Regions , IfI/Plenum, New York. Colorimetric Determinations of Proteins – Conventional methods 1. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry , 1976, 72 :248-54. 2. Read SM and Northcote DH. Minimization of variation in the response to different proteins of the Coomassie blue G dye-binding assay for protein. Analytical Biochemistry , 1981, 116 :53-64. 3. Smith PK; Krohn RI; Hermanson GT; Mallia AK; Gartner FH; Provenzano MD;Fujimoto EK; Goeke NM; Olson BJ; Klenk DC. Measurement of protein using bicinchoninic acid . Analytical Biochemistry , 1985, 150 :76-85. [published erratum appears in Analytical Biochemistry 1987, 163 :279]. Colorimetric Determinations of Proteins – Microplate methods 1. PierceNet.com. Coomassie Plus 2. PierceNet.com. BCA Protein Assay Kit
We have textbook solutions for you!
The document you are viewing contains questions related to this textbook.
Understanding Normal and Clinical Nutrition
The document you are viewing contains questions related to this textbook.
Chapter 22 / Exercise 2
Understanding Normal and Clinical Nutrition
Rolfes/Whitney
Expert Verified
2-9 Copyright © UC Regents Davis campus, 2005-13. All Rights Reserved. May not be re-distributed without prior written consent of course instructor. E XPERIMENTAL A. Q UANTITATIVE PROTEIN ANALYSIS BY ABSORBANCE OF UV LIGHT : A 280 Protein Assay Spectra of amino acids and proteins in the UV Run a spectrum of 1 ml of each solution in quartz cuvettes from 350 - 230 nm, using the Shimadzu spectrophotometer and Spectral Overlay Method (see Page ix of the Introduction to this Manual). compound peak λ (nm) Abs at peak λ Abs at 280 nm 0.80 mg/ml BSA 0.40 mg/ml lysozyme 2.0 mg/ml gelatin A 280 Protein Assay Refer to Table 2.2 (next page) for the A 280 assay protocol to be used. Tubes #1-14 represent the protein standard curve assayed in duplicate using bovine serum albumin (BSA) as a protein standard, while #15-20 are the unknown protein dilutions tested in duplicate. Tubes #21-28 contain other proteins for comparative purposes (sensitivity) and/or substances being tested for interference by absorbing light in the UV range. 1. Obtain the following from the provided stocks solutions: undiluted BSA standard, lysozyme, and gelatin; 2-mercaptoethanol; RNA; SDS; and undiluted unknown protein; in labeled tubes! Also, deionized water. 2. Label 28 microcentrifuge tubes with a Sharpie (#1-28) and place in microcentrifuge test-tube rack. 3. Pipet water, then the interfering substances, and then the proteins into labeled tubes (except for tubes #17-20). Mix each tube gently after the last addition by inverting it two or three times. Do not vortex.

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture