Homework Help is Here – Start Your Trial Now!
Science
Chemistry
B. C. D. E. F. 10nM solution 5nM solution 2.5nM solution 1.25nM solution 0.6nM solution I

B. C. D. E. F. 10nM solution 5nM solution 2.5nM solution 1.25nM solution 0.6nM solution I

Chemistry
Chemistry
10th Edition
ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Chapter1: Chemical Foundations
Section: Chapter Questions
Problem 1RQ: Define and explain the differences between the following terms. a. law and theory b. theory and...
See similar textbooks
icon
Related questions
Question
Convert the amount of spiperone in each sample to the concentration in the 1 ml total volume of assay tube. I have calculated amount of 3H spiperone in pmol in each dilution in second picture. To converted concentration in 1 ml total volume of each assay tube do I need to consider dilution factor？
ABCDELI A. C. D. E. F. G. H. NB. The concentration given above will be the TOTAL spiperone concentration, not necessarily the concentration of 'H-spiperone. The latter needs to be calculated using the SPECIFIC ACTIVITY of the radioligand and the MAX data (see later). Preparation of cell membrane The membrane preparation was diluted in HEPES IA buffer to a concentration of 250μg/ml, so that 100μl of the diluted membrane solution can be placed in each assay tube to give 25µg protein. 20nM solution 10nM solution 5nM solution 2.5nM solution 1.25nM solution 0.6nM solution 0.3nM solution 0.15nM solution Preparation of assay tubes 24 assay tubes were required to measure non-specific binding, and 24 were used to measure total binding of ³H-spiperone to membrane fragments. Each of the 8 spiperone concentrations were assayed in triplicate; 8 concentrations in triplicate requires 24 tubes, made up as follows: 700μl HEPES IA buffer added to 48 assay tubes. ● ● . 100μl of 30μM (+)-butaclamol added to the Non-Specific Binding tubes. 100μl of 30μM (-)-butaclamol added to the Total Binding tubes. 100μl of each [H]-spiperone concentration added to the appropriate assay tubes. ● 100μl of the membrane preparation (25µg) added to each of the assay tubes. Tubes (1.0 ml in total volume) were incubated for I hour in a 25°C waterbath. Harvesting of assay mixtures Membrane preparations were harvested on filter paper under vacuum. The filters, which will have 'trapped' membrane fragments and any ³H-spiperone bound to them, were assayed for '³H using a scintillation counter. 100μl of each ³H-spiperone solution (A-H) were also counted - this formed the MAX data, and represents how much ³H-spiperone was added to each of the tubes. (NB. Remember that this is the same amount of H-spiperone in each dilution that was added to the assay tubes, and therefore will in th FUL Think is yor
Transcribed Image Text:ABCDELI A. C. D. E. F. G. H. NB. The concentration given above will be the TOTAL spiperone concentration, not necessarily the concentration of 'H-spiperone. The latter needs to be calculated using the SPECIFIC ACTIVITY of the radioligand and the MAX data (see later). Preparation of cell membrane The membrane preparation was diluted in HEPES IA buffer to a concentration of 250μg/ml, so that 100μl of the diluted membrane solution can be placed in each assay tube to give 25µg protein. 20nM solution 10nM solution 5nM solution 2.5nM solution 1.25nM solution 0.6nM solution 0.3nM solution 0.15nM solution Preparation of assay tubes 24 assay tubes were required to measure non-specific binding, and 24 were used to measure total binding of ³H-spiperone to membrane fragments. Each of the 8 spiperone concentrations were assayed in triplicate; 8 concentrations in triplicate requires 24 tubes, made up as follows: 700μl HEPES IA buffer added to 48 assay tubes. ● ● . 100μl of 30μM (+)-butaclamol added to the Non-Specific Binding tubes. 100μl of 30μM (-)-butaclamol added to the Total Binding tubes. 100μl of each [H]-spiperone concentration added to the appropriate assay tubes. ● 100μl of the membrane preparation (25µg) added to each of the assay tubes. Tubes (1.0 ml in total volume) were incubated for I hour in a 25°C waterbath. Harvesting of assay mixtures Membrane preparations were harvested on filter paper under vacuum. The filters, which will have 'trapped' membrane fragments and any ³H-spiperone bound to them, were assayed for '³H using a scintillation counter. 100μl of each ³H-spiperone solution (A-H) were also counted - this formed the MAX data, and represents how much ³H-spiperone was added to each of the tubes. (NB. Remember that this is the same amount of H-spiperone in each dilution that was added to the assay tubes, and therefore will in th FUL Think is yor
Table 4 Amount of [H]-spiperone in each assay tube Sample Amount of [³H]- spiperone (pmol) A B C D E F G H 1.35 0.69 0.36 0.19 0.09 0.05 0.02 0.01
Transcribed Image Text:Table 4 Amount of [H]-spiperone in each assay tube Sample Amount of [³H]- spiperone (pmol) A B C D E F G H 1.35 0.69 0.36 0.19 0.09 0.05 0.02 0.01
Expert Solution
trending now

Trending now

This is a popular solution!

steps

Step by step

Solved in 2 steps

SEE SOLUTIONCheck out a sample Q&A here
Blurred answer
Knowledge Booster
Concentration Terms
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, chemistry and related others by exploring similar questions and additional content below.
Similar questions
  • SEE MORE QUESTIONS
Recommended textbooks for you
Chemistry
Chemistry
Chemistry
ISBN:
9781305957404
Author:
Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:
Cengage Learning
Chemistry
Chemistry
Chemistry
ISBN:
9781259911156
Author:
Raymond Chang Dr., Jason Overby Professor
Publisher:
McGraw-Hill Education
Principles of Instrumental Analysis
Principles of Instrumental Analysis
Chemistry
ISBN:
9781305577213
Author:
Douglas A. Skoog, F. James Holler, Stanley R. Crouch
Publisher:
Cengage Learning
Organic Chemistry
Organic Chemistry
Chemistry
ISBN:
9780078021558
Author:
Janice Gorzynski Smith Dr.
Publisher:
McGraw-Hill Education
Chemistry: Principles and Reactions
Chemistry: Principles and Reactions
Chemistry
ISBN:
9781305079373
Author:
William L. Masterton, Cecile N. Hurley
Publisher:
Cengage Learning
Elementary Principles of Chemical Processes, Bind…
Elementary Principles of Chemical Processes, Bind…
Chemistry
ISBN:
9781118431221
Author:
Richard M. Felder, Ronald W. Rousseau, Lisa G. Bullard
Publisher:
WILEY
SEE MORE TEXTBOOKS