Bovine serum albumin, binding with

Figure 11.2 Complexation of Brilliant Green inside CB[7] causes a fluorescence increase which is further enhanced when the complex binds to the protein, bovine serum albumin. Reprinted with permission from [13]. Copyright Wiley-VCH Verlag GmbH Co. KCaA...

Protein concentration can be determined using a method introduced by Bradford,4 which utilises Pierce reagent 23200 (Piece Chemical Company, Rockford, IL, USA) in combination with an acidic Coomassie Brilliant Blue G-250 solution to absorb at 595 nm when the reagent binds to the protein. A 20 mg/1 bovine serum albumin (Piece Chemical Company, Rockford, IL, USA) solution will be used to prepare a standard calibration curve for determination of protein concentration. The sample for analysis of SCP is initially homogenised or vibrated in a sonic system to break down the cell walls. 

The interaction between 4-(4-hydroxybut-2-ynyloxy)-3-(phenylsulfonyl)-l,2,5-oxadiazole-2-oxide 16 and bovine serum albumin (BSA) was studied by spectroscopic methods including fluorescence and UV-Vis absorption spectroscopy. The results indicate that molecules 16 bind with BSA forming 1 1 complex. Thermodynamic parameters, such as AH, AG, and A.Y, were calculated. The results indicate that the binding reaction is mainly entropy driven and hydrophobic forces play a major role in this reaction <2006CHJ1050>. 

Fig. 8.7 CTB-GM1-ganglioside binding ELISA assay. Plates, coated first with GMrganglioside and bovine serum albumin (BSA), respectively, were irrigated with total soluble plant protein from chloroplast transgenic lines (3 and 7) and 300 ng of purified bacterial CTB. The absorbance of the GM1-ganglioside-CTB-antibody complex in each case was measured at 405 nm. Total soluble protein from untransformed plants was used as the negative control.

For quantitative analysis of protein concentration the colorimetric Bradford-assay [147] is most commonly used. Here another Coomassie dye, Brilliant Blue G-250, binds in acidic solutions to basic and aromatic side chains of proteins. Binding is detected via a shift in the absorption maximum of the dye from 465 nm to 595 nm. Mostly calibration is performed with standard proteins like bovine serum albumin (BSA). Due to the varying contents of basic and aromatic side chains in proteins, systematic errors in the quantification of proteins may occur. 

The reactivity of compounds such as 28 was clearly demonstrated by the peroxidase-catalyzed covalent binding of A -methyW-hydroxyellipticine (27) to proteins (756). Using horseradish peroxidase and hydrogen peroxide, tritiated-27 was converted to the 9-oxoellipticine derivative in the presence of bovine serum albumin (BSA) and human antibovine IgG in vitro. Covalent binding to these proteins was confirmed by gel electrophoresis, combustion, and liquid scintillation analysis. Dissolution of the BSA-ellipticinium derivative with pronase and... 

To produce a 1 1 complex with protein (binding to bovine serum albumin)... 

Belgorodsky et al. (2006) studied the binding of pristine fullerene complexed with cyclodextrin on bovine serum albumin protein demonstrating that the binding is a multistep process. First, the cyclodextrin dissociation from C60 takes place, exposing a fullerene hydrophobic portion, than this portion binds to the protein. 

A third issue arises with NCEs that bind to plastic surfaces due to the hydrophobic physical nature of the molecule. A significant loss due to this type of non-specific binding to the Caco-2 device may lead to underestimation of permeability. Addition of protein (e.g., bovine serum albumin) in the receiver compartment appears to be capable of preventing some non-specific binding [23],... 

Fig. 4. Extent of binding of methyl orange at pH 7.0 and 25°C as a function of free (nonbound) dye concentrations (1) polyethylenimine with 8.4% of residues acylated by lauroyl groups (2) polyethylenimine with 11.5% of residues acylated by hexanoyl groups (3) polyethylenimine with 10% of residues acylated by butyrl (O) or isobutyrl ( ) groups (4) Polyethylenimine, PEI-600 (5) bovine serum albumin.

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