In addition to our purified DHFR sample, we will also be including three other protein samples as controls. Of the different protein samples analyzed, which protein will travel the slowest/least distance through the SDS-PAGE gel? A) The smallest protein analyzed B) The largest protein analyzed C) Although different in size, all proteins analyzed will travel the same distance on the gel

1a) In MALDI-TOF data, when performing peptide mass mapping search,  the number of peptides and the sequence coverage decreases, but protien score increase. Explain b) As the same as part a, the peptide masses and the coverage remain the same, but the protien score increase, explain how this can happen.

4) Researcher #1 and #2 in your lab each conducted a protein purification. They present you with the following data. The protein-of-interest is indicated by the red arrowhead, and the numbers on top indicate the purification step. Researcher #1 1 2 3 4 SDS-PAGE (Coomassie Stained) Researcher #2 1 2 3 4 5 6 Immunoblot a) Which researcher has more clearly demonstrated purification of the protein-of-interest? Justify your answer. b) What type of chromatography step did researcher #1 most likely conduct in going from step 2➜ 3 based on the gel? Justify your answer.

What does -log(p-value) tell in immunoprecipitation mass spectrometry? What does log2 Fold change say? Generally, what does the graph of immunoprecipitation mass spectrometry tell (ie. how to read its graph)?

In Western Blots and ELISAS, the following molecules are used:  Tween-20 SDS Goat anti-rabbit IgG (whole molecule) – HRP Rabbit anti-egg albumin Bovine IgG Rabbit anti-bovine IgG – HRP Egg albumin (globular protein – what is Molecular weight?) Milk protein (globular protein – what is Molecular weight?) TMB Beta-mercaptoethanol Methanol HCl May you explain the reasons why they are used?