What's important in this lab?
- What property is electrophoresis using?
- How will we determine if Rubisco is present? Can we?
- What are the molecular weights of the 2 Rubisco polypeptide subunits?
- What travels fastest/farthest? Small proteins or large proteins?
So we've used solubility and charge to isolate Rubisco from Spinach leaves. What's left?
SDS Page uses a gel to run the proteins (Filtrate, P1, P1(low/med/high), P2, P2(low/med/high), and marker).
Why is the marker important?
- We know the molecular weights of the 2 Rubisco sub-units (55,000 and 14,000). The marker will allow us to know the molecular weights of the proteins in the pellets and supernatants.
Did you find Rubisco?? Compare your results to the marker! (Notice, other proteins are still in the mix, see why we did all these steps?!)
Smaller proteins travel faster and farther! They end up at the bottom of the gel! Large proteins are too fat to move, they move sloooowww!
Here's a way to remember this: (compliments of Dr. Poole)
I'm a girl, and if I do say so myself I am a thin girl! My friends on the other hand, are 6ft 5in, 250lb men! HUGE! When we go out, they can see over the crowd, but getting to the bar is another story, they are toooo big!
Not going to lie, I want a drink when I'm there and I'm not willing to wait! So my little self can weave in and out of the crowd and make my way to the bar! DRINKS FOR ALL! I win because I am small, I can get through the crowd faster than my 250lb friends, all I need them to do is grab their drinks over everyone else's head!
So technically, I am the protein in the gel. I am smaller so I can get to the finish faster, I get to the end (the bar) before everyone else!
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