BCA assay 정량

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• 1. BCA assay

  BCA assay는 단백질 정량 방법 중 하나로, 구리 이온이 단백질과 반응하여 발색하는 원리를 이용합니다. A 용액과 B 용액을 50:1 비율로 섞어 사용하며, 표준 단백질 농도를 0, 0.125, 0.25, 0.5, 1, 2 ug/ml로 준비하여 흡광도를 측정합니다. 이를 통해 샘플의 단백질 농도를 계산할 수 있습니다. 샘플 버퍼, 단백질 양, 용해 버퍼의 양 등을 고려하여 western blotting 실험에 사용할 수 있습니다.

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• 1. BCA assay

  The BCA (Bicinchoninic Acid) assay is a widely used method for quantifying the total protein concentration in a sample. It is a colorimetric detection technique that relies on the reduction of copper ions (Cu2+) to cuprous ions (Cu+) by proteins in an alkaline medium, followed by the chelation of the cuprous ions with bicinchoninic acid to produce a purple-colored complex. This complex exhibits a strong absorbance at 562 nm, which can be measured using a spectrophotometer to determine the protein concentration. The BCA assay is a popular choice for protein quantification due to its high sensitivity, wide linear range, and compatibility with a variety of sample types, including cell lysates, tissue homogenates, and purified protein solutions. It is also relatively simple to perform, making it a versatile and accessible technique for researchers in various fields, such as biochemistry, molecular biology, and cell biology. One of the key advantages of the BCA assay is its ability to tolerate the presence of common interfering substances, such as detergents, reducing agents, and salts, which can often interfere with other protein quantification methods like the Bradford assay. This makes the BCA assay particularly useful for analyzing samples that contain these components, such as those obtained from cell culture or protein purification experiments. Additionally, the BCA assay is compatible with a wide range of protein concentrations, typically ranging from 20 μg/mL to 2,000 μg/mL, allowing researchers to accurately quantify both low and high protein concentrations in their samples. This flexibility makes the BCA assay a valuable tool for a variety of applications, including enzyme activity assays, protein-protein interaction studies, and the optimization of protein purification protocols. Overall, the BCA assay is a robust and reliable method for protein quantification that has become a staple in many biochemical and molecular biology laboratories. Its versatility, sensitivity, and tolerance to common interfering substances make it a valuable technique for researchers working with a wide range of protein-containing samples.