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So if a protein A interacts with protein B but we need to know if it is subunit B1 or B2, can we use the same method by doing two pull down assays ( one with protein A and B1 and one with protein A and B2)? Or is there another technique for that?
Thank you! However, in future videos, maybe it would be better if you could add some possible controls for the technique. e.g. having only beads as a control to identify non-specific binding.
Your videos are really very interesting and beneficial especially your Drosophila related. Are u working on Drosophila? as during my summer training I watched your videos for basics.
Hiii everybody ! I made a pull down essay but I dont understand how I am suppose to present the result and how I have to read it. In the video its explain really quickly in the end in 2:10 min. Someone can explain to me ? I made input WB, Pull down WB and Pull down Coomassie
Make a correction in your video bro: GST is not part of plasmids, plasmids contain genes of interest, not proteins of interest. Genes of interest then encode proteins, here the GST works as bait. GST is added, not encoded by the plasmids as you have shown in your diagram.
GST is a 211 amino acid protein (26 kDa) whose DNA sequence is frequently integrated into expression vectors for production of recombinant proteins. The result of expression from this vector is a GST-tagged fusion protein in which the functional GST protein (26 kDa) is fused to the N-terminus of the recombinant protein. Because GST folds rapidly into a stable and highly soluble protein upon translation, inclusion of the GST tag often promotes greater expression and solubility of recombinant proteins than expression without the tag. In addition, GST-tagged fusion proteins can be purified or detected based on the ability of GST (an enzyme) to bind its substrate, glutathione (GSH).
@@animatedbiologywitharpan agreed bro, still, you need to use word ''GST,s DNA sequence'' as you you mentioned in your reply. But in video, maybe unconsciously, you used the word GST Protein in plasmid. As you know the sequence will translate into GST protein.
