Pull on your thinking caps this dark and wet morning, it's time for another addition of "Science so easy even a monkey could do it."
*Insert cool bumper that shows lots of B&W footage of monkeys in lab coats playing with flasks and beakers*
I like to mess with people by saying that I'm working on H1N1 (you may remember, not so deadly swine flu that the mass media got worked into a froth over. Hm, maybe it was dangerous in that it caused the media to turn rabid. May have to look into that), and I have been but it's just been a simple ELISA plate. So no reason to put the Hazmat suits on just yet.
ELISA stands for Enzyme Linked something something Assay. Okay so the name isn't really important, but what it does is.
Let's say you have a specific protein you're looking for. You want to find out just how much this little guy has appearasatwigase:One of the easiest approaches is to use an ELISA plate. What it does is that the amount of protein inside each plate well that contains one sample correlates to a color change. More protein = more blue. How it works is a bit more complicated.
You start out with a 96 well plate (the backbone of any micro lab) that is coated with an anti-protein to the protein your looking for. It works like a lego piece, the anti has just the right holes so that the protein your looking for will fit right in. You then add a conjugate that adheres to your protein and has a special color indicator. So that when a chemical is added it will change colors and light up if present.
Actually we're gonna need some visuals, okay here we go. This should clear things right up:The red present represents the anti-protien that is coated in every well. It's just sitting there waiting for its protein soul mate to come along. Groucho represents the protein we're looking for (in this case the dreaded Swine Flu). It fits perfectly with the red present, while the skull and cross bones which is not our protein (we can call it media rabies) does not attach to the red present.
The Cigar is the conjugate. It attaches easily to Groucho (it is Groucho after all) but will not adhere to Mr. Skull Head. Now you just add a chemical and the cigar shows a blue color in the left where Grouch --our wanted protein-- is present while it turns no colors with Mr. Skull head because he never adhered and formed the link.
You then take your plate and put it in a reader that will determine just how much blue is contained in your plate and using a few calculations you can determine just how much of your protein is present.Aside from the two controls, as you can see, we didn't have very much H1N1 though we didn't expect much either.
And that is how you perform an ELISA. It's a rather simple and straight forward idea which means it's a giant pain to get to work perfectly in lab and likes to fail a lot of the time. In a lab anything described as simple will take you three days to get working right. Anything described as complicated will take you a lifetime.
I hope you all enjoyed another round of "Science made easy." Next week we shall tackle the mysteries of how to get a microscope to focus on something other than the back of your eye and just what does make the sky blue (hint, it involves elves with a paint brush).
Same lab channel, same lab time.