Analysis of a 49kDA protein is aimed with a western blot technique. For this purpose, "whole cell extract" was obtained from biopsy samples taken from three different individuals, and when their spectrophotometric measurements were made, their concentrations were determined to be 40 µg/µL, 50 µg/µL, and 100 µg/µL for three samples, respectively. At the beginning of the experiment, 200 µg of protein is loaded into each gel well by using a ready 5x loading solution containing SDS and β-mercapta-ethanol together with glycerol and methylene blue dye. a) If loading will be done in a volume of 20µL, then how is the preparation of the samples before loading? b) Considering the loading content given above, what can be said about whether this PAGE is "non-denaturing" or "denaturing". Justify taking into account the loading solution content.
Analysis of a 49kDA protein is aimed with a western blot technique. For this purpose, "whole cell extract" was obtained from biopsy samples taken from three different individuals, and when their spectrophotometric measurements were made, their concentrations were determined to be 40 µg/µL, 50 µg/µL, and 100 µg/µL for three samples, respectively. At the beginning of the experiment, 200 µg of protein is loaded into each gel well by using a ready 5x loading solution containing SDS and β-mercapta-ethanol together with glycerol and methylene blue dye. a) If loading will be done in a volume of 20µL, then how is the preparation of the samples before loading? b) Considering the loading content given above, what can be said about whether this PAGE is "non-denaturing" or "denaturing". Justify taking into account the loading solution content.
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Analysis of a 49kDA protein is aimed with a western blot technique. For this purpose, "whole cell extract" was obtained from biopsy samples taken from three different individuals, and when their spectrophotometric measurements were made, their concentrations were determined to be 40 µg/µL, 50 µg/µL, and 100 µg/µL for three samples, respectively. At the beginning of the experiment, 200 µg of protein is loaded into each gel well by using a ready 5x loading solution containing SDS and β-mercapta-ethanol together with glycerol and methylene blue dye.
a) If loading will be done in a volume of 20µL, then how is the preparation of the samples before loading?
b) Considering the loading content given above, what can be said about whether this PAGE is "non-denaturing" or "denaturing". Justify taking into account the loading solution content.
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