You are studying the toxic protein called ectatomin, a major component of ant venom (specifically from the species Ectatomma tuberculatum), that embeds into cell membranes and creates pores that cause cells to lyse. Ectatomin is a small dimer, meaning the protein is comprised of two polypeptides; the sequences of the two polypeptides are given in the table with single-letter abbreviations. polypeptide sequences of ectatomin monomers polypeptide protein sequence polypeptide 1 GVIPKKIWETVCPTVEPWAKKCSGDIATYIKRECGKL polypeptide 2 WSTIVKLTICPTLKSMAKKCEGSIATMIKKKCDK You could use absorbance at 280?? or colorimetric assays like the Bradford assay or bicinchoninic acid assay (BCA) assay to quantify the amount of ectatomin you have for your experiment. Why might you choose to use a colorimetric method instead of absorbance at 280??? Select the best answer. Measuring absorbance at higher wavelengths, like 595?? or 562??, is more accurate than measuring absorbance at 280??. The Bradford and BCA assays require less time to setup and measure protein concentration. Ectotamin has few tryptophan and tyrosine residues, so the absorbance at 280?? will be too low to accurately measure unless the sample is very concentrated. The Coomassie dye or the bicinchoninic acid in the Bradford or BCA assay will react with tryptophan and tyrosine residues to give a stronger absorbance reading, making it easier to measure the concentration of samples with low amounts of protein.

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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  1. You are studying the toxic protein called ectatomin, a major component of ant venom (specifically from the species Ectatomma tuberculatum), that embeds into cell membranes and creates pores that cause cells to lyse. Ectatomin is a small dimer, meaning the protein is comprised of two polypeptides; the sequences of the two polypeptides are given in the table with single-letter abbreviations.

polypeptide sequences of ectatomin monomers

polypeptide

protein sequence

polypeptide 1

GVIPKKIWETVCPTVEPWAKKCSGDIATYIKRECGKL

polypeptide 2

WSTIVKLTICPTLKSMAKKCEGSIATMIKKKCDK

You could use absorbance at 280?? or colorimetric assays like the Bradford assay or bicinchoninic acid assay (BCA) assay to quantify the amount of ectatomin you have for your experiment.

Why might you choose to use a colorimetric method instead of absorbance at 280??? Select the best answer.

  1. Measuring absorbance at higher wavelengths, like 595?? or 562??, is more accurate than measuring absorbance at 280??.
  2. The Bradford and BCA assays require less time to setup and measure protein concentration.
  3. Ectotamin has few tryptophan and tyrosine residues, so the absorbance at 280?? will be too low to accurately measure unless the sample is very concentrated.
  4. The Coomassie dye or the bicinchoninic acid in the Bradford or BCA assay will react with tryptophan and tyrosine residues to give a stronger absorbance reading, making it easier to measure the concentration of samples with low amounts of protein.
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