28 Dec The technique that needs to have a

The technique that needs to have an experimental design and a drawn figure is IMMUNOHISTOCHEMISTRY.

1) Experimental design – describe the experiment you are going to do. Be sure to include what elements of the system described below that experiment will prove (each individual experiment is not likely to prove everything… for example you may only show that two proteins interact, not that they are required for localization – or vice versa – they are required for localization, but you have not demonstrated that direct interaction). Also be sure to describe the variables of your experiment – so for example, if you are going to use an antibody be sure to describe which protein that antibody binds to. Be sure to include any controls that your experiment might require to confirm that your answer is correct.

-Purpose: The main goal of the experiment is to test whether nfkb moves from the cytoplasm to the nucleus.

-Basic experimental design including what the variables of your experiment are…example : are you stimulating cells to activate NFKB?

– Controls ?

2) Figure – draw the figure that you would expect if your results prove the system described below. Be sure to include labels on the figure, and any information that you need to interpret the figure. NOTE: if you are drawing an image of cells it is standard to include a nuclear dye so that it is clear where the individual cells are. (Just drawing a bunch of dots in a square does not constitute a good figure.) For some good advice on this look up published figures that have used this technique. Many companies will describe a technique and give you a gallery of images that have used this technique (generally with their antibodies that they are trying to sell you) these are probably particularly nice figures.

Must have the following elements

1) Labels – is the figure labels appropriately?

2) Images consistent with the experiment

3) Control images

4) Images that are interpretable (not just dots or lines on a page)

The Scenario: You are studying a transcription factor NFkappaB. This transcription factor has a nuclear localization sequence (NLS) as a part of its amino acid sequence. When NFkappaB is bound to the protein IkappaB that NLS is covered and the two proteins are retained in the cytoplasm. However, when IkappaB is ubiquitinated and degraded then the NLS on NFkappaB is exposed and NFkappaB interacts with the Nuclear Pore proteins and is moved into the nucleus where it increases the production of certain genes.

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