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Bradford Ultra快速考马斯亮蓝法测定蛋白含量
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4度
12个月
wuhao
大量
上海武昊公司
1L
BradfordUltra 已配制好可直接使用的快速考马斯亮蓝法测定蛋白含量的试剂盒
Expedeon 的 Bradford Ultra 是已配制好可直接使用的快速考马斯亮蓝法测定蛋白含量的试剂盒,环境中可含有高达1%的去垢剂(1%高蛋白质范围, 0.1%低蛋白质范围)。 当考马斯亮蓝染料在酸性介质结合蛋白后, 吸收最大值立即发生转变,从465nm变为595nm,相应颜色从棕色变成蓝色。其光吸收值与蛋白质含量成正比,因此可用于蛋白质的定量测定。 Expedeon的BradfordUltra是对传统的Bradford配方的改进,传统的Bradford 方法不能承受大于0.1%的去垢剂的蛋白质样品。此外, 与传统的Bradford 方法相比,BradfordUltra试剂有更好的线性范围和更少的protein-to-protein变化。 优点 –快速(5 - 10分钟) – 操作简单 –还原剂兼容 –去垢剂兼容
Comparison of Expedeon's BradfordUltra Assay with Pierce's "Coomassie Protein Assay. The graph shows the average blank corrected A595 measurements for detergents samples (no protein). When detergents are present in the sample, competitor Bradford assay solutions give high blank readings making accurate detection of the protein sample impossible. BradfordUltra is not affected by the detergents compared to Classic formulations and provides excellent signal to noise ratios and accurate determintation of the protein concentration regardless of the detergent presence.
Standard curves obtained with BradfordUltra are unaffected by the presence of detergents.
Standard curves obtained with classical Bradford formulation are significantly affected by thepresence of detergents resulting in loss of sensitivity and inaccurate results.
1) Make a dilution series of the chosen model protein in the range:
0.1 mg/ml - 1.5 mg/ml (high protein range) OR
1 g/ml - 25 g/ml (low protein range)
2) Mix the samples, standards and a blank (buffer, no protein) with BradfordUltra reagent.
High Protein Range: 1 part sample for 15 parts reagent
Low Protein Range: 1 part sample for 1 part reagent
3) Read absorbance at 595 nm.
4) Calculate concentration:
Subtract the average 595 nm measurement for the blank from the 595 nm measurements of all other individual standards and unknown samples. Plot the average blank-corrected 595 nm measurement for each standard vs. concentration. Use the slope of this standard curve to estimate the protein concentration of the unknown samples.
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2020.12.28-2021.01.31
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