Quantitation of proteins using a dye–metal-based colorimetric protein assay

Antharavally, Babu S.; Mallia, Krishna A; Rangaraj, Priya; Haney, Paul; Bell, Peter A.

doi:10.1016/j.ab.2008.11.024

Cited by 50 publications

(34 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results of protein quantitation with ethanolic extracts of Ixora coccinea, Hibiscus rosa-sinensis, Lawsonia inermis, Jatropha curcas and Hibiscus sadarifa are quite promising. The sensitivity is comparable to the well-known methods of Lowry et al [2], Bradford [3] or Pierce 660 nm protein assay of Antharavally et al [5]. Every protein assay has limitations which essentially depends on the specific protein sample being analyzed, composition of the solution containing the protein and the type of sensitivity required.…”

Section: Discussionmentioning

confidence: 50%

See 1 more Smart Citation

Protein quantitation using dyes obtained from plant materials

Akinseye¹,

Odutayo²,

Adewale³

2012

ABB

View full text Add to dashboard Cite

We present a preliminary report on the use of plant dyes in the quantitation of proteins in solution. We have used ethanol, acid, alkali and water to extract dyes from some plant materials, including flowers of Jungle flame (Izora coccinea), China rose (Hibiscus rosa-sinensis) and leaves of West African Indigo (Lonchocarpus cyanescens), Mimosa (Mimosa pudica), Roselle (Hibiscus sabdarifa), Jatropha (Jatropha curcas) and Henna (Lawsonia inermis). The dyes obtained were used in the protein-dye binding studies. The colour of the protein-dye complex of the ethanolic extracts was stable and increased linearly with increase in protein concentration. The extracts achieved linearity up to the following amount of proteins in the test samples: Hibiscus rosa-sinensis (60 g), Ixora coccinea (120 g), Hibiscus sabdarifa (80 -100 g), Jatropha curcas (80 g), and Lawsonia inermis (100 g). The sensitivity of the dyes especially at low protein concentrations indicate that they can provide suitable alternatives to other well known standard methods of protein determination.

show abstract

Section: Discussionmentioning

confidence: 50%

“…The methods of bicinchoninic acid (BCA) and Pierce 660 nm protein assay [4,5] are based on protein-metaldye-binding assay. Again, users who have relied on these methods have brought to the fore, some problems associated with this procedure [6].…”

Section: Introductionmentioning

confidence: 99%

Protein quantitation using dyes obtained from plant materials

Akinseye¹,

Odutayo²,

Adewale³

2012

ABB

View full text Add to dashboard Cite

show abstract

“…The amount of SOD protein was determined by a dye-metal-based colorimetric protein assay [72]. The assay was conducted in a microplate format and a commercially available reagent (Pierce 660 nm Protein Assay Reagent) was used.…”

Section: Methodsmentioning

confidence: 99%

Superoxide Dismutase as a Novel Macromolecular Nitric Oxide Carrier: Preparation and Characterization

Chen

Chiu

2012

IJMS

View full text Add to dashboard Cite

Nitric oxide (NO) is an important molecule that exerts multiple functions in biological systems. Because of the short-lived nature of NO, S-nitrosothiols (RSNOs) are believed to act as stable NO carriers. Recently, sulfhydryl (SH) containing macromolecules have been shown to be promising NO carriers. In the present study, we aimed to synthesize and characterize a potential NO carrier based on bovine Cu,Zn-superoxide dismutase (bSOD). To prepare S-nitrosated bSOD, the protein was incubated with S-nitrosoglutathione (GSNO) under varied experimental conditions. The results show that significant S-nitrosation of bSOD occurred only at high temperature (50 °C) for prolonged incubation time (>2 h). S-nitrosation efficiency increased with reaction time and reached a plateau at ~4 h. The maximum amount of NO loaded was determined to be about 0.6 mol SNO/mol protein (~30% loading efficiency). The enzymatic activity of bSOD, however, decreased with reaction time. Our data further indicate that NO functionality can only be measured in the presence of extremely high concentrations of Hg2+ or when the protein was denatured by guanidine. Moreover, mildly acidic pH was shown to favor S-nitrosation of bSOD. A model based on unfolding and refolding of bSOD during preparation was proposed to possibly explain our observation.

show abstract

“…Protein quantification is performed using the Coomassie-based Bradford protein assay [36,37]. When comparing method A with method B (n = 6), more proteins were extracted when the membrane was removed after centrifugation (method A).…”

Section: Methodsmentioning

confidence: 99%

Practical Problems Encountered during the Cultivation of an Open-Source Reconstructed Human Epidermis Model on a Polycarbonate Membrane and Protein Quantification

Lemper

Paepe²,

Rogiers³

2013

Skin Pharmacol Physiol

View full text Add to dashboard Cite

During recent years, the importance of in vitro technology in skin research has increased significantly. A variety of skin culture models have been developed and commercialized. In this respect, the availability of reconstructed human epidermis (RHE) equivalents represents a significant improvement compared to the use of monolayer cultures. However, when an in-house RHE model is being developed, researchers might encounter some difficulties during cultivation. The scope of this paper is to report our experiences and practical problems with the development of a three-dimensional RHE model cultured on a polycarbonate membrane. Some important issues including cell density, the use of lysing enzymes, culture media, cell storage and viability, cell confluency and protein extraction are reported and optional solutions are given.

show abstract

Quantitation of proteins using a dye–metal-based colorimetric protein assay

Cited by 50 publications

References 9 publications

Protein quantitation using dyes obtained from plant materials

Protein quantitation using dyes obtained from plant materials

Superoxide Dismutase as a Novel Macromolecular Nitric Oxide Carrier: Preparation and Characterization

Practical Problems Encountered during the Cultivation of an Open-Source Reconstructed Human Epidermis Model on a Polycarbonate Membrane and Protein Quantification

Contact Info

Product

Resources

About