[유전학실험]Exp 4. Protein extraction and Quantitative analysis

소개글

Exp 4. Protein extraction and Quantitative analysis

목차

Ⅱ. Introduction
Ⅰ. Abstract
Ⅲ. Material
IV. Chemical reagent & Apparatus

본문내용

2) Protein quantitation: Bradford assay
The Bradford assay, a colorimetric protein assay, is based on an absorbance shift in the dye Coomassie when bound to arginine and hydrophobic amino acid residues present in protein.
The bound form of the dye is blue and has an absorbance spectrum maximum held to be at 595 nm. The unbound forms are green(650 nm) and red(470 nm). The increase of absorbance at 595 nm is proportional to the amount of bound dye, and thus to the amount(concentration) of protein present in the sample.
Unlike other protein assays, the Bradford protein assay is less susceptible to interference by various chemicals that may be present in protein samples.
Bradford assay Standard Curve
장점
-빠르고 (10분)저렴하다.
-단백질에 매우 특이적이다.(Highly specific)
-높은 강도를 가진다.(sensitive)
-다양한 단백질에 이용 가능하다.
-dye-complex는 약 1시간 동안 안정하다.
단점
-넓은 범위에 걸친 Standard Curve의 경우 non-linear 하다.
-각각의 단백질에 대한 반응이 다양할 수 있으므로 Standard의 선택이 중요하다.
-사용되는 단백질은 비가역적으로 변성된다.

참고 자료

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