How to calculate using the protein markers, estimate the number and sizes of pro
ID: 176469 • Letter: H
Question
How to calculate using the protein markers, estimate the number and sizes of protein molecules in EACH sample.
Sample Type
Concentration determined from Bradford Assay (3pts)
Volume of sample extract needed for approx. 40 µg (6pts)
Laemmli Buffer*
Deionized water needed for 40µL total volume (this can be zero if your sample is very dilute) (6pts)
Egg Whites
13.3 µg/µL
10 µL
26.5 µL
Fish Muscle
22.6 µg/µL
10 µL
Chicken Muscle
21.5 µg/µL
10 µL
Sample Type
Concentration determined from Bradford Assay (3pts)
Volume of sample extract needed for approx. 40 µg (6pts)
Laemmli Buffer*
Deionized water needed for 40µL total volume (this can be zero if your sample is very dilute) (6pts)
Egg Whites
13.3 µg/µL
3.5ml
10 µL
26.5 µL
Fish Muscle
22.6 µg/µL
5.5ml10 µL
24.5µLChicken Muscle
21.5 µg/µL
5.0ml10 µL
25µLExplanation / Answer
We estimate number and sizes of protein molecules by running them into sodium dodecyl sulfate used Polyacrylamide gel electrophoresis (SDS -PAGE) method. Protein molecules are in tertiary or quaternary structure, so we use SDS for linearizing the protein molecules. We make protein sample by boiling into SDS and after it add loading dye to load sample into well and color dye for protein staining. We load sample into the wells made by a comb placed over Acrylamide- Bisacrylamide gel. In this gel we also add markers of protein which make bands when run. These bands are observed with corresponded sample band and then find the size of protein. Number of bands tells the number of proteins in a sample. Buffers are used when we run the sample; all wells and gel should be embedded into buffers. Buffers allow running of sample by providing electrophoretic movement.
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