In-gel total protein quantification using a ninhydrin-based method

Amino Acids ◽  
2013 ◽  
Vol 45 (4) ◽  
pp. 1003-1013
Author(s):  
Sung Ung Kang ◽  
Seok Heo ◽  
Gert Lubec
Keyword(s):  
Total Protein ◽  
Protein Quantification

scholarly journals Heat and Pressure Treatments on Almond Protein Stability and Change in Immunoreactivity after Simulated Human Digestion

Nutrients ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 1679 ◽  
Author(s):  
Elisabetta De Angelis ◽  
Simona Bavaro ◽  
Graziana Forte ◽  
Rosa Pilolli ◽  
Linda Monaci
Keyword(s):  
Protein Stability ◽  
Total Protein ◽  
Protein Pattern ◽  
Protein Quantification ◽  
Protein Profiling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Total Protein Content ◽  
Bioinformatic Tools ◽  
Sds Page ◽  
Allergenic Proteins

Almond is consumed worldwide and renowned as a valuable healthy food. Despite this, it is also a potent source of allergenic proteins that can trigger several mild to life-threatening immunoreactions. Food processing proved to alter biochemical characteristics of proteins, thus affecting the respective allergenicity. In this paper, we investigated the effect of autoclaving, preceded or not by a hydration step, on the biochemical and immunological properties of almond proteins. Any variation in the stability and immunoreactivity of almond proteins extracted from the treated materials were evaluated by total protein quantification, Enzyme Linked Immunosorbent Assay (ELISA), and protein profiling by electrophoresis-based separation (SDS-PAGE). The sole autoclaving applied was found to weakly affect almond protein stability, despite what was observed when hydration preceded autoclaving, which resulted in a loss of approximately 70% of total protein content compared to untreated samples, and a remarkable reduction of the final immunoreactivity. The final SDS-PAGE protein pattern recorded for hydrated and autoclaved almonds disclosed significant changes. In addition, the same samples were further submitted to human-simulated gastro-intestinal (GI) digestion to evaluate potential changes induced by these processing methods on allergen digestibility. Digestion products were identified by High Pressure Liquid Chromatography-High Resolution Tandem Mass Spectrometry (HPLC-HRMS/MS) analysis followed by software-based data mining, and complementary information was provided by analyzing the proteolytic fragments lower than 6 kDa in size. The autoclave-based treatment was found not to alter the allergen digestibility, whereas an increased susceptibility to proteolytic action of digestive enzymes was observed in almonds subjected to autoclaving of prehydrated almond kernels. Finally, the residual immunoreactivity of the GI-resistant peptides was in-silico investigated by bioinformatic tools. Results obtained confirm that by adopting both approaches, no epitopes associated with known allergens survived, thus demonstrating the potential effectiveness of these treatments to reduce almond allergenicity.

Quantification of Protein Solutions with Trinitrobenzenesulfonic Acid

1970 ◽  
Vol 16 (1) ◽  
pp. 24-31 ◽  
Author(s):  
Jesse F Goodwin ◽  
Siu-Ying Choi
Keyword(s):  
Cerebrospinal Fluid ◽  
Spectrophotometric Method ◽  
Total Protein ◽  
Protein Quantification ◽  
Protein Solutions ◽  
Complexation Reaction ◽  
Trinitrobenzenesulfonic Acid ◽  
Factors Affecting ◽  
Spectral Absorbance ◽  
Protein Components

Abstract Trinitrobenzenesulfonic acid (TNBS) may be used for quantifying dilute solutions of protein. The trinitrophenyl protein—sulfite complexes resulting from reaction of TNBS with protein in the presence of sulfite are highly colored and exhibit maximum spectral absorbance at 420 nm. The TNBS protein—sulfite complex is formed by a simple two-stage condensation and complexation reaction. Protein in concentrations as low as 20 µg may be detected by the reaction. The method devised for protein quantification compares favorably with an ultraviolet spectrophotometric method and the conventional biuret procedure. Bilirubin and hemoglobin interference with the TNBS method is negligible. The procedure is useful for measuring total protein and globulin in serum and cerebrospinal fluid and also for measuring minor serum protein components. Procedural parameters and factors affecting the measurement of protein by the TNBS—sulfite reaction are presented.

scholarly journals Quantitative Method for Assessing the Role of Lysine & Arginine Post-Translational Modifications in Nonalcoholic Steatohepatitis

2020 ◽  
Author(s):  
Aaron E. Robinson ◽  
Aleksandra Binek ◽  
Vidya Venkatraman ◽  
Brian C. Searle ◽  
Ronald J. Holewinski ◽  
...  
Keyword(s):  
Total Protein ◽  
Protein Quantification ◽  
Regulatory Function ◽  
The Novel ◽  
Localization Algorithm ◽  
Post Translational Modifications ◽  
Simultaneous Quantification ◽  
Data Independent Acquisition ◽  
Site Localization

AbstractProteoforms containing post-translational modifications (PTMs) represent a degree of functional diversity only harnessed through analytically precise simultaneous quantification of multiple PTMs. Here we present a method to accurately differentiate an unmodified peptide from its PTM-containing counterpart through data-independent acquisition-mass spectrometry, leveraging small precursor mass windows to physically separate modified peptidoforms from each other during MS2 acquisition. We utilize a lysine and arginine PTM-enriched peptide assay library and site localization algorithm to simultaneously localize and quantify seven PTMs including mono-, di-, and tri-methylation, acetylation, and succinylation in addition to total protein quantification in a single MS run without the need to enrich experimental samples. To evaluate biological relevance, this method was applied to liver lysate from differentially methylated non-alcoholic steatohepatitis (NASH) mouse models. We report altered methylation and acetylation together with total protein changes drive the novel hypothesis of a regulatory function of PTMs in protein synthesis and mRNA stability in NASH.Graphical Abstract

P08.19 Comparative evaluation of surgical cycle performed in washer-disinfectors by visual inspection versus total protein quantification

2010 ◽  
Vol 76 ◽  
pp. S27-S28
Author(s):  
V. Sebastián ◽  
B. Peláez ◽  
L. Barreales ◽  
R. Andrade ◽  
C. Fernández ◽  
...  
Keyword(s):  
Total Protein ◽  
Comparative Evaluation ◽  
Visual Inspection ◽  
Protein Quantification

scholarly journals Total protein level reduction of odontopathogens biofilms by Rosmarinus officinalis L. (rosemary) extract: an analysis on Candida albicans and Streptococcus mutans

2019 ◽  
Vol 22 (2) ◽  
pp. 260-266
Author(s):  
Jonatas Rafael De Oliveira ◽  
Gabriela de Fátima Santana-Melo ◽  
Samira Esteves Afonso Camargo ◽  
Luana Marotta Reis De Vasconcellos ◽  
Luciane Dias De Oliveira
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Total Protein ◽  
Protein Level ◽  
Protein Composition ◽  
Rosmarinus Officinalis ◽  
Protein Quantification ◽  
Rosemary Extract ◽  
Total Protein Level ◽  
Rosmarinus Officinalis L

Objective: The resistance of fungi and bacteria to the available antimicrobials has increased and the development of alternative products to control them has become a very requirement. The use of plant products could be a viable option due to the efficacy, viability, and availability they present. Thereby, this study evaluated the effect of R. officinalis L. extract on C. albicans and S. mutans biofilms, by the total protein level analysis presented by the microorganisms. Material and Methods: For this purpose, monomicrobial biofilms were formed for 48 h and exposed to the R. officinalis L. extract for 5 min. Then, total protein quantification was performed by Lowry method. Results: The analysis showed significant total protein reduction of the biofilms after exposure to the plant extract, with 39 ± 11%, for C. albicans, and 32 ± 11%, for S. mutans. Conclusion: R. officinalis L. extract decreased the total protein level in both biofilms. Thus, C. albicans and S. mutans protein composition could be a target for action of antimicrobial agents.KeywordsBiofilms; Candida albicans; Proteins; Rosmarinus officinalis; Streptococcus mutans.

scholarly journals Investigation of Heat and Pressure Treatments on Almond Protein Stability and Immunoreactivity after Simulated Human Digestion

2018 ◽  
Author(s):  
Elisabetta De Angelis ◽  
Simona L. Bavaro ◽  
Graziana Forte ◽  
Rosa Pilolli ◽  
Linda Monaci
Keyword(s):  
Total Protein ◽  
Protein Pattern ◽  
Protein Quantification ◽  
Protein Profiling ◽  
Total Protein Content ◽  
Bioinformatic Tools ◽  
Sds Page ◽  
Allergenic Proteins ◽  
The Stability

Almond is worldwide consumed and renowned as a valuable healthy food. In spite of this, it is also a potent source of allergenic proteins able to trigger several mild to life-threatening immunoreactions. Food processing proved to alter biochemical characteristics of proteins, thus affecting the respective allergenicity. In this paper we investigated the effect of autoclaving, preceded or not by a hydration step, on the biochemical and immunological properties of almond proteins. Any variation in the stability and immunoreactivity of almond proteins extracted from the treated materials, were evaluated by total protein quantification, ELISA assay and protein profiling by electrophoresis-based separation (SDS-PAGE). The autoclaving alone was found to weakly affect almond proteins stability, despite what observed for the combination of hydration and autoclaving, which resulted in a loss of approximately 70% of total protein content compared to untreated sample, and in a final negligible immunoreactivity, as well. The final SDS-PAGE protein pattern recorded for almonds hydrated and autoclaved disclosed significant changes. In addition, the same samples were further submitted to in vitro simulated gastro-duodenal (GI) digestion to evaluate potential changes induced by these processing on allergens digestibility. Digestion products were identified by HPLC-HRMS/MS analysis followed by software-based data mining, and complementary information were provided by analyzing the proteolytic fragments lower that 6 kDa in size. The autoclave based treatment was found not to alter the allergens digestibility, whereas an increased susceptibility to proteolytic action of digestive enzymes was observed in almonds subjected to the combination of prehydration and autoclaving. Finally, the residual immunoreactivity of the GI resistant peptides was investigated in-silico by bioinformatic tools, confirming that by following both approaches, no epitopes survived the almond digestion, thus demonstrating the potential effectiveness of these treatments to reduce almond allergenicity.

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