An improved bradford protein assay for collagen proteins

López, JoséManuel; Imperial, Santıago; Valderrama, Rodrigo; Navarro, Salvador

doi:10.1016/0009-8981(93)90009-s

Cited by 54 publications

(20 citation statements)

References 20 publications

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“…Co, USA) and the samples were centrifuged at 2.5 x 10 3 rpm, for 20 minutes at 4 C. The protein content was determined by the Micro-Bradford assay as usual. 21,30 The quantitative results were mea-sured by a microphotometer (Dynatech MR7000, USA) and were given as a concentration of optical density (OD 490) over protein mass. Samples of 1mg/ml protein concentration were stored at -20°C.…”

Section: Preparation Ofprotein Fractions Containing Aquaporinsmentioning

confidence: 99%

Development and characterisation of a monoclonal antibody family against aquaporin 1 (AQP1) and aquaporin 4 (AQP4)

Nagy

Szekeres²,

Kvell

et al. 2002

Pathol. Oncol. Res.

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Recent studies have discovered the existence of water-channel molecules, the so called aquaporins (AQP) presumably involved in active, ATP dependent water transport between the intracellular and extracellular compartments. Both genetic and protein sequences and structures of the AQPs are known and crystallographic analyses of some members of the AQP family have been performed. Specific antibodies are required to examine their histological locations and analyse their roles in physiological and pathological pathways of water transportation and osmotic regulation. Until recently some polyclonal antibodies have been developed against certain members of the AQP family. However, to date highly specific monoclonal antibodies against aquaporins do not exist. We have developed a monoclonal antibody family against the aquaporin 1 (AQP1) and aquaporin 4 (AQP4) molecules. Well-conserved epitop sequences of AQP1 and AQP4 proteins were selected by computer analysis and their synthetic peptide fragments were used as the antigens of immunisation and the following screening. Antibodies were characterised by immunoserological methods (ELISA, dot-blot and immunoblot), flow cytometry and immunohistochemistry of formaldehyde-fixed and paraffin-embedded tissue samples. RT-PCR tests controlled the specificity of the immune reactions. Our monoclonal antibodies recognised AQP1 and AQP4 in all the techniques mentioned above and apparently they are useful both in various quantitative and qualitative measurements of the expressions of AQP1 and AQP4 in several species (human, rat, mouse, invertebrates, even plants). According to preliminary immunohistochemical studies our monoclonal anti-AQP1 and anti-AQP4 antibodies are appropriate tools of patho-morphological examinations on routine formol-paraffin tissue samples.

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Section: Preparation Ofprotein Fractions Containing Aquaporinsmentioning

confidence: 99%

Development and characterisation of a monoclonal antibody family against aquaporin 1 (AQP1) and aquaporin 4 (AQP4)

Nagy

Szekeres²,

Kvell

et al. 2002

Pathol. Oncol. Res.

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“…Proteins contain basic amino acid groups such as arginine, histidine, and lysine in their structure. For BSA, 16.5 % of its groups are basic amino acids [24]. Therefore, BSA forms a blue dye-BSA complex that can be detected by a UV-Vis spectrophotometer at 595 nm [22].…”

Section: Resultsmentioning

confidence: 99%

An investigation of folic acid–protein association sites and the effect of this association on folic acid self-assembly

et al. 2015

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The contribution of folic acid (FA)-tryptophan interactions to FA-protein association was investigated in the context of using FA as a drug carrier in protein delivery systems. Bovine serum albumin (BSA) and indolicidin were used as model proteins in the study. The FA-BSA complex was characterized by using the Bradford reagent to identify the impact of FA-BSA association on BSA-dye reagent interactions. UV-visible spectroscopic analysis of the FA-BSA mixture showed that the absorbance maximum of BSA-dye reagent occurred at 595 nm, even after the association of FA with BSA. This confirms that protonated amino acid groups of the protein are not involved in FA-BSA association. Moreover, molecular dynamics (MD) simulation confirmed the presence of an associative interaction between aromatic moieties in FA and tryptophan moieties in the indolicidin molecule, which disrupted FA self-assembly. An X-ray diffraction (XRD) study showed that there was limited disruption of FA self-assembly after the addition of BSA or tryptophan. This suggests that FA and BSA are compatible and associate with each other. Graphical Abstract Mechanism of folic acid and protein association.

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“…According to the high porosity and high surface area of the obtained PCL mats, the coating process was done via dipping the mats into a polyP/collagen solution as follows: the collagen suspension (10 mL), containing 0.1 g of collagen (determined according to the Bradford method [35,36,37]; Shenzhen Lando Biomaterials, Shenzhen, China) was diluted to 50 mL in distilled water. Then 0.1 g of Na-polyP was added and the reaction solution was kept at pH 10.…”

Section: Methodsmentioning

confidence: 99%

Morphogenetically-Active Barrier Membrane for Guided Bone Regeneration, Based on Amorphous Polyphosphate

et al. 2017

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We describe a novel regeneratively-active barrier membrane which consists of a durable electrospun poly(ε-caprolactone) (PCL) net covered with a morphogenetically-active biohybrid material composed of collagen and inorganic polyphosphate (polyP). The patch-like fibrous collagen structures are decorated with small amorphous polyP nanoparticles (50 nm) formed by precipitation of this energy-rich and enzyme-degradable (alkaline phosphatase) polymer in the presence of calcium ions. The fabricated PCL-polyP/collagen hybrid mats are characterized by advantageous biomechanical properties, such as enhanced flexibility and stretchability with almost unaltered tensile strength of the PCL net. The polyP/collagen material promotes the attachment and increases the viability/metabolic activity of human mesenchymal stem cells compared to cells grown on non-coated mats. The gene expression studies revealed that cells, growing onto polyP/collagen coated mats show a significantly (two-fold) higher upregulation of the steady-state-expression of the angiopoietin-2 gene used as an early marker for wound healing than cells cultivated onto non-coated mats. Based on our results we propose that amorphous polyP, stabilized onto a collagen matrix, might be a promising component of functionally-active barrier membranes for guided tissue regeneration in medicine and dentistry.

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An improved bradford protein assay for collagen proteins

Cited by 54 publications

References 20 publications

Development and characterisation of a monoclonal antibody family against aquaporin 1 (AQP1) and aquaporin 4 (AQP4)

Development and characterisation of a monoclonal antibody family against aquaporin 1 (AQP1) and aquaporin 4 (AQP4)

An investigation of folic acid–protein association sites and the effect of this association on folic acid self-assembly

Morphogenetically-Active Barrier Membrane for Guided Bone Regeneration, Based on Amorphous Polyphosphate

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