BCH 3346 Final: BCH3346 Final Exam Study Guide

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blackbutterfly828

30 Mar 2016

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Learning objectives: explain why the bradford assay must be preformed under acidic condition, compare and contrast three forms of cbb-g250 that can exist. The bradford assay is a dye-based spectrophotometric assay which allows for the determination of protein concentration in an aqueous sample. Essentially, this means that a colour change is produced when protein-dye complex forms, this change in colour of the dye can be detected as a shift in wavelength, measured by a spectrophotometer. Because it is the anionic form (blue) that non-covalently binds to protein, the equilibrium shifts from the cationic form (red) to the anionic from (blue) as bbg binds to proteins. Thus, the cationic form acts as a reservoir of the binding dye. The reaction is preformed under acidic conditions (ph < 1) so that bbg is mostly in the cationic, form that absorbs at. The two other forms are also present, although in lower proportion.

Related Questions

Q1.

During the process of solubilisation of proteins, Dithiothreitol and Î²-Marcaptoethanol are used in order to:

Select ONE answer:

A. Reduce the disulphide bonds present between and within protein chains

B. Act as denaturing agents

C. Both a and b

D. None of these

Q2.

High amount of salts is present in many biological samples. They are generally removed by:

Select ONE answer:

A. Spin dialysis and Dialysis

B. Gel filtration

C. Buffer exchange

D. All of the above

Q3.

Nucleic acids are generally removed during the sample preparation of proteins. Which of the following statement is NOT correct for nucleic acid contaminants?

Select ONE answer:

A. They can bind to proteins through electrostatic interactions

B. They enhance the sample viscosity and cause background smearing

C. Higher molecular weight nucleic acid can clog gel pores

D. They donât interfere with proteins in gels because of their higher molecular weight

Q4.

During protein sample preparation by TRIzol method, the sample is divided into three different layers. What of the following is correct for those layers?

Select ONE answer:

A. Pink â DNA, White -Protein, Transparent-RNA

B. Pink - Protein, White - DNA, Transparent-RNA

C. Pink- RNA, White - protein, Transparent- DNA

D. Pink- Protein, White- RNA, Transparent- DNA

Q5.

Bradford assay is commonly used to quantify the protein concentration in a protein sample. Which of the statements given below is NOT correct for the Bradford assay?

Select ONE answer:

A. The assay is based on the absorbance shift of Coommassie brilliant blue dye.

B. Red form of the dye is converted into its blue form under acidic conditions.

C. Absorbance is always measured at 280 nm.

D. Bradford assay is based on Beer Lambert law.

BCH 3346 Final: BCH3346 Final Exam Study Guide

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