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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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Book ChapterDOI
TL;DR: This chapter describes the purification of several Rabkinesin-6 domains from bacteria and eukaryotic systems, and their biochemical characterization (ATPase, MT-binding, hydrodynamic properties) and an assay to visualize the interaction between Rab6A and Rabkine-6.
Abstract: This chapter describes the purification of several Rabkinesin-6 domains from bacteria and eukaryotic systems, and their biochemical characterization (ATPase, MT-binding, hydrodynamic properties)The chapter describes an assay to visualize the interaction between Rab6A and Rabkinesin-6 For purification, Escherichia coli strain freshly transformed with pET-Nt or pTrc-Q665 plasmids are cultured Bacterial pellets are obtained are sonicated, centrifuged, and chromatographed Protein fractions are analyzed by SDS-PAGE and Western blot This protocol leads to the purification of about 05–1 mg fusion proteins α/β-Tubulin is purified from bovine brain with two cycles of polymerization in the presence of GTP, and microtubule (MT)-associated proteins (MAPs) are removed by phosphocellulose chromatography Tubulin concentration is estimated by the Bradford assay and MT concentration is expressed per tubulin heterodimer MT-induced ATPase activity of kinesins is often measured by using a NADH coupled enzymatic assay

12 citations

Journal ArticleDOI
TL;DR: An inexpensive and simple application of the Bradford assay is developed to determine the residual protein content (RPC) in cell culture supernatants and its performance is more robust and accurate over time and the respective concentration range.
Abstract: Frequently measured mammalian cell culture process indicators include viability and total cell concentration (TCC). Cell lysis, an additional important process characteristic that substantially contributes to the overall product purity profiles, is often not addressed in detail. In the present study, an inexpensive and simple application of the Bradford assay is developed to determine the residual protein content (RPC) in cell culture supernatants. The reliability and reproducibility of the method are tested in a long-term study and compared with lysis quantification via the DNA measurement. The results show that its performance is more robust and accurate over time and the respective concentration range. Additionally, both methods are used for cell lysis process monitoring in a recombinant Chinese hamster ovary fed-batch process. In the presented process, by applying the established assay, the lysis rate k DL is determined to be constant over time at 4.6 × 10 -4 lysed cell concentration (LCC) per TCC and time (LCC/TCC/h). In contrast, DNA data did not confirm the constant lysis rate due to variations of the content per cell during cultivation. Thus, information on the RPC can facilitate the determination of the optimal harvest time point with respect to purity and in improving process characterization.

11 citations

Journal ArticleDOI
TL;DR: Large inconsistencies in replicating anaerobic runs for protein concentration appeared to be explained by noting that rising CO 2 bubbles may cause ‘foam fractionation’ of the proteins in the broth.

11 citations

Journal ArticleDOI
TL;DR: In this article, field natural rubber latex was irradiated with different doses near a 60Co gamma source to reduce the water-soluble protein content in the final product, the protein content of the films obtained by casting method was extracted with phosphate buffer solution, pH 7 and was measured using Micro BCA Protein Assay kit.

11 citations

Journal ArticleDOI
01 Jul 2011
TL;DR: The methods considered in this lecture have multi-year history and applied widely in the laboratory practice, there are some crucial points, which must be taken into consideration while choosing the method permitting reliably and with a high specificity and reproducibility to quantify protein this paper.
Abstract: Protein quantification is an integral part of any investigation related to protein isolation, purification, characterization, and analysis. Although the methods considered in this lecture have multi-year history and applied widely in the laboratory practice, there are some crucial points, which must be taken into consideration while choosing the method permitting reliably and with a high specificity and reproducibility to quantify protein.

11 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
20238
202212
202127
202021
201919
201822