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en:quantification_of_protein [2013/06/09 10:10] (current)
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 +======Quantification of Protein======
 +There are two major principles to quantify concentration of proteins.
 +タンパク質の定量法には、大きくわけて二つの方法がある。
 +  -Spectrophotometric method
 +  -Colorimetric method - relative quantification using some standard substances such as [[BSA]] or [[casein]].
 +
 +
 +
 +^                   ​^ ​ Type              ^  Detection wavelength (nm)  ^  Approx. sensitivity ​  |
 +^ABS (near-UV) ​     |Spectr. ​ |  280 nm     ​| ​ 20-3000 µg  |
 +^Lowry method ​      ​|Color. ​  ​| ​ 770 nm     ​| ​  5-100 µg   |
 +^ABS (near-UV) ​     |Spectr. ​ |  205 nm     ​| ​  1-100 µg   |
 +^Bradford method ​   |Color. ​  ​| ​ 595 nm     ​| ​  1-50 µg    |
 +^BCA method ​        ​|Color. ​  ​| ​ 562 nm     ​| ​ 0.2-50µg ​   |
 +
 +
 +=====Spectrophotometric method=====
 +{{icon>​flask_large}}
 +
 +**Extinction coefficient**\\ \\ 
 +**ε<​sub>​280nm</​sub>​** (M<​sup>​-1</​sup>​cm<​sup>​-1</​sup>​) = 5500N<​sub>​Tyr</​sub>​+1490N<​sub>​Trp</​sub>​+125N<​sub>​Cys</​sub>​\\ \\ 
 +**[protein]** = A<​sub>​280nm</​sub>​ x ε<​sub>​280nm</​sub>​ M<​sup>​-1</​sup>​\\ \\ 
 +N<​sub>​Tyr</​sub>,​N<​sub>​Trp</​sub>,​N<​sub>​Cys</​sub>:​Amino acids' count
 +
 +
 +======References======
 +
 +  *{{pubmed>​long:​19892168}}
 +  *{{pubmed>​long:​18265371}}
 +  *{{pubmed>​long:​10075906}}
 +  *{{pubmed>​long:​8563639}}
 +
 +
 +======Link======
 +
 +  *[[:​タンパク質の定量法]]
 +
 +
 +{{tag>​en:​Proteins en:​Quantification}}