Draw the Western Blot that would result from undertaking the same experiment in the presence of: i) An estrogen receptor agonist ii) An estrogen receptor antagonist iii) An estrogen receptor agonist in the presence of an excess amount of antagonist iv) Estrogen in the presence of an excess amount of antagonist v) An estrogen receptor inverse agonist.

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Chapter1: The Human Body: An Orientation
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Draw the Western Blot that would result from undertaking the same experiment in the presence of:

i) An estrogen receptor agonist

ii) An estrogen receptor antagonist

iii) An estrogen receptor agonist in the presence of an excess amount of antagonist

iv) Estrogen in the presence of an excess amount of antagonist

v) An estrogen receptor inverse agonist. 

This question relates to the biological activity of the estrogen receptor (ER). ER is a
nuclear receptor and positively regulates the expression the c-Myc gene, a
transcription factor found at elevated levels in numerous cancers.
To understand the effect of drugs on the biological behaviour of the estrogen receptor
in cells, you undertake Western Blotting analysis of a cell line that expresses a
recombinant human estrogen receptor. After culturing the cells, you then lyse the cells
and fractionate them, generating membrane, cytosolic and nuclear protein fractions
from the whole cell lysate. After running these fractions on SDS-PAGE gels, you
transfer to nitrocellulose membrane and probe with monoclonal antibody conjugates
for the proteins ER and c-Myc.
The results of your Western Blotting experiment without any treatment are provided in
Figure 1.
Primary antibody
Cell fraction
Anti-ER
Anti-c-myc
Membrane
Cytoplasm
Nucleus
| |
Transcribed Image Text:This question relates to the biological activity of the estrogen receptor (ER). ER is a nuclear receptor and positively regulates the expression the c-Myc gene, a transcription factor found at elevated levels in numerous cancers. To understand the effect of drugs on the biological behaviour of the estrogen receptor in cells, you undertake Western Blotting analysis of a cell line that expresses a recombinant human estrogen receptor. After culturing the cells, you then lyse the cells and fractionate them, generating membrane, cytosolic and nuclear protein fractions from the whole cell lysate. After running these fractions on SDS-PAGE gels, you transfer to nitrocellulose membrane and probe with monoclonal antibody conjugates for the proteins ER and c-Myc. The results of your Western Blotting experiment without any treatment are provided in Figure 1. Primary antibody Cell fraction Anti-ER Anti-c-myc Membrane Cytoplasm Nucleus | |
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