Chem 162 (Biochemistry Lab) - Exam #1
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What do we measure at 280 nm?
Proteins with aromatic side chains
What do we measure at 260 nm?
Nucleic acid presence
What do we measure at 190 nm?
Absorbance by peptide bond presence
What do we measure at 340 nm?
- Absorbance by NADH
- >> NAD+ cannot absorb light at this range
What do we measure at 595 nm?
- The amount of bound Coomassie Blue (amt of Coomassie Blue that has bound to protein). This thus is used to give us an idea of what the [Protein Concentration] is.
What charge does Sulfonate have? What is it used in?
- - Sulfonate has a negative charge
- - Sulfonate is the anion (stationary phase) used in Cation Exchange Chromatography
What are the requirements for a buffer?
- - The pKa's that you use must be within +/- 1 unit of the pH that we're using (or making), because if not then the equilibrium for recreating our buffer (conjugate acid/base) will be thrown off
- - There must be a sufficient (high enough) concentration of our buffer
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