Colorimetric Assays Flashcards
1
Q
What do colorimetric assays aim to do?
A
Determine the concentration of substances based on their color change
2
Q
What is the device used to measure the amount of light absorbed?
A
Spectrophotometer
3
Q
What are the 2 main colorimetric assays?
A
The Bradford assay and ELISA
4
Q
Benefits of colorimetric assay?
A
- Simple, easy, and relatively quick
- Cost-effective
- Quite versatile
- Mostly non-destructive
5
Q
What are the downsides of colorimetric assays?
A
- Low sensitivety
- Lack of Specificity (if interference from similar compounds cause same color)
- Human error, calibration error
6
Q
Why would you use an assay?
A
- Overall protein concentration of a sample
- Identify specific protein in sample
- Also used for enzymatic activity
7
Q
What is the dye in the bradford assay?
A
Coomassie Brilliant Blue
8
Q
What is the color change of Coomassie?
A
Brown if no protein
Blue if protein
darker = more
9
Q
3 main parts to a bradford assay?
A
- Making the standard curve
- Preparing the tissue
- Analysis
10
Q
What is used to make standard curve and why?
A
BSA, bc cheap, and often has no effect in many biochemical reaction
11
Q
Why does a new standard curve have to be done each time?
A
- consistency in conditions
- Minimises degradation
12
Q
A
