Hello, I'm new here (obviously).
I'm a Psychology major, with a BMS minor, attending Central Connecticut State University. I figured that if I wanted to help remember some of the jargon in relation to my minor, it would greatly benefit me to emerse myself into any kind of science environment--such as these forums! I'm a Junior at the moment, however I have found some of my sophomore level classes can be quite trying--such as BMS 201.
Right now the topic we're covering is Bradford Assays (since we're studying proteins, and their varying structures / roles). I have a lab report to write on it coming up (tomorrow actually), but I want to make sure that I have my science down, before I go writing verbal diarrhea and making a fool of myself. Since I don't have many science friends, this is pretty much my only option (aside from several other forums, but for now I'll start with just the one).
My concerns are over what exactly the purpose of the Bradford Reagent is, and why it is used--likewise, why BSA is used / what the purpose of it is. We used the BSA to establish a standard curve, using 0microL (control), 2microL, 4microL, 6microL, and 8microL of the BSA in microtiter plates (done twice to get a comparative average).
Then, after establishing a standard curve we took 3microL of 5 different foods (Rice Milk, Coffee Creamer, Soy Sauce, Chili & Sour Cream). We picked these randomly from 20+ different food variants.
So to bring us up to speed:
200microL of the Bradford reagent was used within each of the 20 microtiter wells. Well rows A-B contained the 0-8microL of BSA in well #s 1-5. Well rows A-B, #s 6-10 contained the food stuffs. Then the microtiter plate was read by a spectrophotometer at an OD of 595nm.
I will provide the data in hopes of getting the best help in analyzing this information, and ensuring I understand the analysis.
BSA OD Readout
0microL - 0.00
2microL - .281
4microL - .538
6microL - .748
8microL - .880
Food Stuffs OD Readout (3microL each) BEFORE dividing by 3, in order of highest OD readout, to lowest.
1.) Creamer - 1.1505 (0g protein per 2g serving) = 0.00% protein content (5)
2.) Sour Cream - .913 (1g protein per 28.4g serving) = 3.52% protein content (3)
3.) Rice Milk - .189 (.4g protein per 245g serving) = .163% protein content (4)
4.) Chili - .121 (17g protein per 227g serving) = 7.49% protein content (2)
5.) Soy Sauce - .104 (2g protein per 18g serving) = 11.1% protein content (1)
(The hyphens are dividers, not minus signs)
When looking up the ingredients I was surprised to find that Cremora (the brand of Coffee Creamer used) had no protein content what-so-ever, and while I was expecting Chili to be a close runner for 1st or 2nd highest protein concentration, it was near dead last--this, I am assuming, is due to when the Chili was cooked, the proteins had been denatured? Yet that still doesn't explain why Cremora scored so high (although to get a 1 mg/ml BSA solution ratio we divided the food stuffs results by 3). Which then leads to the next question, what is the difference between denatured proteins and naturally occurring proteins? (Not a Lab question, but just a personal one) For example: why is cooked chicken good for promoting muscle building, as opposed to it raw (aside from obvious food-borne illnesses), but also, why wouldn't it (denatured proteins) be useful for this assay?
When looking at the data as a whole, it almost goes perfectly inverse in relationship to what common sense--given the protein contents imply--as far as readouts are concerned.
Thanks for reading, I hope I was concise enough in what I am asking! Feel free to leave any feedback, commentary, etc.