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WorksheetsWestern Blotting
Total questions: 7
Worksheet time: 3mins
After proteins are run on an SDS-PAGE gel, a transfer is executed. What is the purpose of the transfer in Western blot protocol?
Denature the proteins in the sample
Visualize the proteins run on the gel
Move proteins from the SDS-PAGE gel to a nitrocellulose membrane
Probe the gel with an antibody to detect a protein of interest
Why is it called immunoblott?
because it involves human antibody
because the membrane contains antibody
because it applies fluorescent antibodies
because it applies antibody to tag protein
because the protein of interest is an antibody
Which is the right orientation?
cathode - filter paper - gel - transfer membrane - filter paper - anode
cathode - filter paper - transfer membrane - gel - filter paper - anode
What is the purpose of washing in every step in western blotting?
to ensure a stable pH environment in the tank
to avoid protein degradation
to remove any unbound antibodies
to moisten the transfer membrane
to remove any contaminating residues
State the purpose of blocking
to block nonspecific antibody from binding to protein
to avoid non specific binding of antibody
to avoid antibodies from binding to protein
to prevent protein from being washed off
to prevent unwanted contamination towards proteins
Primary antibody targets..
nitrocellulose surface
specific protein
primary antibody
conjugated secondary antibody
fluorochrome
Conjugated secondary antibody targets..
nitrocellulose surface
specific protein
primary antibody
conjugated secondary antibody
fluorochrome
