Freimoser:Protocols:ProteinQuantification
From OpenWetWare
Jump to navigationJump to search
<html><style type='text/css'> A:link {
text-decoration: underline; font-size: 10pt; font-style: normal; font-family: arial, trebuchet ms; color: #003399;
}
A:visited {
text-decoration: underline; font-size: 10pt; font-style: normal; font-family: arial, trebuchet ms; color: #003399;
} A:active {
text-decoration: none; background-color: #99ccff; color: #660099;
}
A:hover {
text-decoration: none; color: #ff0033;
}
.navbar{
padding: 0px; width: "100%"; font-family: Arial; font-size: 10pt;
} </style></html>
Source
The quantification methods listed below represent those that we find most useful. The descriptions are part of a more detailed list of protein quantification methods [1]
General Reference: Stoscheck, CM. Quantitation of Protein. Methods in Enzymology 182: 50-69 (1990). [2]
Absorbance assays
Absorbance at 280 nm
- Range: 20 μg to 3 mg
- Volume: Depends on cuvette - volumes range from 200 microliters to 3 ml or greater
- Accuracy: Fair
- Convenience: Excellent, if equipment available
- Major interfering agents: Detergents, nucleic acids, particulates, lipid droplets
Absorbance at 205 nm
- Range: Roughly 1 to 100 μg
- Volume: Depends on cuvette - volumes range from 200 microliters to 3 ml or greater
- Accuracy: Fair
- Convenience: Very good
- Major interfering agents: Detergents, nucleic acids, particulates, lipid droplets
Colorimetric assays
Modified Lowry
- Range: 2 to 100 μg
- Volume: 1 ml (scale up for larger cuvettes)
- Accuracy: Good
- Convenience: Fair
- Major interfering agents: Strong acids, ammonium sulfate
Bradford assay
- Range: 1 to 20 μg (micro assay); 20 to 200 μg (macro assay)
- Volume: 1 ml (micro); 5.5 ml (macro)
- Accuracy: Good
- Convenience: Excellent
- Major interfering agents: None
