After running your purified DHFR fusion protein+Laemmli 40uL sample, along with a protein ladder you transfer protein on the gel
to a nitrocellulose filter for Western Blot analysis. a. Which of the following primary antibodies could you use to detect your fusion protein based on the fusion protein identified and process described above: Anti-Myc, Anti-FLAG, Anti-GST, Anti-His, Anti-DHFR, Anti-Tubulin? List all that apply
Anti-his and Anti-DHFR antibodies can be used to detect the fusion as the primary antibodies in the process.
Explanation:
Western blot is often used in research to separate and identify proteins. Two major processes are involved in Western blot, namely, SDS-polyacrylamide gel electrophoresis and protein blotting and testing.
In the process above, the DHFR fusion protein was purified and run in the SDS-PAGE ( sds- page separates protein based on molecular weight) for the western blotting. This DHFR protein was fused with histadine tag which is why the antibody (anti-his) can be used as a primary antibody (Primary antibodies directly bind to the protein of interest). Anti DHFR can also be used as a primary antibody.
The DHFR protein was fused with His tag. The DHFR fusion protein was purified and run in the SDS-PAGE for the western blotting. After the completion of gel run, the protein was transferred into the nitrocellulose membrane.
a. For the detection of fusion protein two types of primary antibody can be used. These are Anti-his and Anti-DHFR.
Anti-his antibody can be used as the protein was tagged with histidine, and anti-DHFR antibody can also be used.
