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Bradford protein assay
About: Bradford protein assay is a research topic. Over the lifetime, 635 publications have been published within this topic receiving 239107 citations.
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TL;DR: The CV method was especially suitable for assaying large numbers of samples and was shown to measure the cytotoxicity in several compounds in a reproducible and precise manner.
Abstract: Summary Simplified methods were described for the measurement of growth of human carcinoma cells in tissue culture: ( a ) determination of color intensity in lysates of cells stained with crystal violet (CV method), ( b ) determination of the reduction of 2,6-dichlorophenol indophenol, and ( c ) determination of total organic solids (dichromate method). The three methods gave comparable measurements of relative cell growth and correlated well with the more laborious protein assay. The CV method was especially suitable for assaying large numbers of samples and was shown to measure the cytotoxicity in several compounds in a reproducible and precise manner. Standard errors per assay were comparable between protein and CV assays (approximately 20 per cent).
33 citations
10 Sep 2015
TL;DR: In this paper, the authors used antigen B of hydatid cyst fluid as a sample and the assay was performed in microplate wells and calculated the protein concentration of sample was calculated using the equation of the standard curve.
Abstract: Background: Bradford protein assay is popular due to its ease of performance and relative sensitivity. Many researchers and laboratories in Iran use standard assay of Bradford by cuvette. No commercial kit was available for Bradford microplate assay in Iran. Meanwhile, imported Bradford commercial kits are very expensive and have a long delivery time in Iran. Till now no study or document on Bradford microplate protein quantification assay was reported in Iran, so this study aimed to design and carried out this assay. Methods: In the current study, antigen B of hydatid cyst fluid was used as sample and the assay was performed in microplate wells. The absorbance values were measured at 595 nm and standard curve was generated by Microsoft Office Excel software. The protein concentration of sample was calculated using the equation of the standard curve. Results: Average protein concentration of the sample was 1175 ;mug/ml. The total time needed for reading of absorbance was two minutes approximately. Conclusion: Bradford microplate protein assay is a fast and suitable method. This method could be replacing the time consuming method with cuvette. In addition, if this assay produces as a low price kit it could have many benefits for students and laboratories that need to determine protein concentration by Bradford assay.
33 citations
TL;DR: A modified version of the Coomassie Brilliant Blue dye binding protein assay has been developed for oceanographic samples and intercalibrated with the widely used Lowry assay as discussed by the authors.
33 citations
TL;DR: The aim of this manuscript was to study the application of a new method of protein quantification in Candida antarctica lipase B commercial solutions and calculated error in protein content ranged from 400 to 1200% when protein in CALB solution was quantified.
33 citations
TL;DR: This chapter describes a procedure using Tar-containing membranes and purified soluble chemotaxis proteins that demonstrates a role for the Tar chemoreceptor and the CheW protein in modulating the phosphorylation reactions.
Abstract: Publisher Summary This chapter describes a procedure using Tar-containing membranes and purified soluble chemotaxis proteins that demonstrates a role for the Tar chemoreceptor and the CheW protein in modulating the phosphorylation reactions. Additionally, reference to heterologous systems using chimeras made by fusing portions of Tar to other receptors proteins for testing their effect on phosphate transfer reactions and/or signaling are presented for comparison. For protein assay, the total protein concentration of each preparation is measured using the Bio-Rad protein reagent concentrate, with bovine serum albumin as a standard. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is performed by using 12.5% slab resolving gels. The position of proteins on the gel is determined by Coomassie staining or Western blot analysis. CheW is expressed from the plasmid pDV4 in strain KO685. For purification of CheW, strain KO685 with the pDV4 plasmid is cultivated in LB containing 100 μg/ml ampicillin at 37 ° until logarithmic phase. Tar-containing membranes are isolated from a strain containing a plasmid which overexpresses the protein using a tac promotor P. Plasmids expressing mutant Tar proteins are constructed by replacement of a restriction fragment containing the wild-type sequence with a corresponding mutant fragment. In this way, both "tumble" and "smooth" receptor-encoding vectors are constructed. These mutant Tar proteins proved useful in verification of the specificity of the activation of phosphorylation. All plasmids are maintained in a strain deleted of all four Escherichia coli chemoreceptors. This strain is also used as a source of negative control membranes.
33 citations