Total protein level measurement in bronchoalveolar lavage fluid (BALF) of mice measurement using the Bradford Protein Assay Kit from OZ Biosciences

Total protein levels in BALF of mice were measured using a Bradford Protein Assay Kit from OZ Biosciences.

This article demonstrate the capacity of the Bradford Protein Assay Kit from OZ Biosciences  to efficiently measure the total protein concentration in BALF of mice.

article reference: Shock. 2014 Mar 21. 

TLR2 Deficiency Aggravates Lung Injury Caused by Mechanical Ventilation.

Kuipers MT, Jongsma G, Hegeman MA, Tuip-de Boer AM, Wolthuis EK, Choi G, Bresser P, van der Poll T, Schultz MJ, Wieland CW.

Abstract

Innate immunity pathways are found to play an important role in ventilator-induced lung injury. We analyzed pulmonary expression of Toll-like receptor 2 (TLR2) in humans and mice and determined the role of TLR2 in the pathogenesis of VILI in mice.TLR2 gene expression was analyzed in human bronchoalveolar lavage fluid (BALF) cells and murine lung tissue after 5 hours of ventilation. In addition, wild type (WT) and TLR2 knockout (KO) mice were ventilated with either lower tidal volumes (VT) of 7ml/kg with positive end-expiratory pressure (PEEP), or higher VT of 15ml/kg without PEEP for 5 hours. Spontaneously breathing mice served as controls. Total protein and immunoglobulin M (IgM) levels in BALF, neutrophil influx into the alveolar compartment, and interleukin (IL)-6, IL-1β and keratinocyte-derived chemokine (KC) concentrations in lung tissue homogenates were measured. We observed enhanced TLR2 gene expression in BALF cells of ventilated patients and in lung tissue of ventilated mice. In WT mice, ventilation with higher VT without PEEP resulted in lung injury and inflammation with higher IgM levels, neutrophil influx and levels of inflammatory mediators compared to controls. In TLR2 KO mice neutrophil influx and IL-6, IL-1β and KC were enhanced by this ventilation strategy. Ventilation with lower VT with PEEP only increased neutrophil influx and was similar in WT and TLR2 KO mice. In summary, injurious ventilation enhances TLR2 expression in lungs. TLR2 deficiency does not protect lungs from VILI. In contrast, ventilation with higher VT without PEEP aggravates inflammation in TLR2 knockout mice.

The Bradford Protein Assay Kit (B-Pak) from OZ Biosciences is a straightforward and rapid procedure for determining the concentration of protein in solution.The B-Pak is based on the binding of Coomassie Brilliant Blue G-250 dye to the proteins and particularly basic and aromatic amino acids residues. The dye exists in three forms: cationic (red), neutral (green) and anionic (blue).