Answered: As Build's laboratory partner, help him…

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

Section: Chapter Questions

Problem 1P

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Question

As Build's laboratory partner, help him determine the following:
1. maximum amount of the unknown protein (in mL) that must be loaded into the column
2. elution volume (in mL) of the proteins in the calibration mixture
3. elution volume (in mL) of the unknown protein
4. linear regression parameters (slope, y-intercept, r) of the standard curve used to identify th
molecular weight of the unknown protein (report the exact values)
5. molecular weight (in kDa) of the unknown protein (report in the nearest whole number)
6. most probable identity of the unknown protein based on the table below
PROTEIN
OFF
GUN
BIBLE
OHM
NANON
Page 5 of 7
MOLECULAR WEIGHT, kDa
23
44
76
93
116

$You and Build Jakapan were tasked to determine the molecular weight of an unknown protein using gel filtration chromatography. You made use of a Sephadex G-100 column with a height of 30.0 cm and an internal volume of 52.0 mL. Initially, blue dextran was made to run through the column, giving an elution volume of 3.0 mL. Next, a calibration mixture containing phytochrome 730 (124 kDa), hemoglobin (64.5 kDa), and horse myoglobin (17 kDa) was made to flow into the column, generating a flow rate of 2.0 mL per minute. Shown as follows is the illustration of the collected fractions. Fraction number: ¯ÐÐ¯¯¤¤ÐÐ¶¶ÐÐ ÐÐ¡¡8 1 2 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Subsequently, a sample of the unknown protein was loaded into the column. The elution profile of the unknown protein is shown below. Absorbance at 254 nm 0.6 0.5 0.4 0.3 0.2 0.1 0 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Elution Volume, mL$

Transcribed Image Text:You and Build Jakapan were tasked to determine the molecular weight of an unknown protein using gel filtration chromatography. You made use of a Sephadex G-100 column with a height of 30.0 cm and an internal volume of 52.0 mL. Initially, blue dextran was made to run through the column, giving an elution volume of 3.0 mL. Next, a calibration mixture containing phytochrome 730 (124 kDa), hemoglobin (64.5 kDa), and horse myoglobin (17 kDa) was made to flow into the column, generating a flow rate of 2.0 mL per minute. Shown as follows is the illustration of the collected fractions. Fraction number: ¯ÐÐ¯¯¤¤ÐÐ¶¶ÐÐ ÐÐ¡¡8 1 2 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Subsequently, a sample of the unknown protein was loaded into the column. The elution profile of the unknown protein is shown below. Absorbance at 254 nm 0.6 0.5 0.4 0.3 0.2 0.1 0 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Elution Volume, mL

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