Binding bovine serum albumin

Carbamazepine-protein binding Bovine serum albumin Dialysis (for sampling) Fluorimetry 6 x 10-6 mol L 1 Flow injection system in vitro study of the binding mechanism of the analyte with albumin mini-column of Pb02(s) for oxidising the analyte to a fluorescent product [528]... 

This experiment provides a nice example of the application of spectroscopy to biochemistry. After presenting the basic theory for the spectroscopic treatment of protein-ligand interactions, a procedure for characterizing the binding of methyl orange to bovine serum albumin is described. 

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. 

Specifications for chromatographic packings often describe maximum loading in terms of the maximum capacity of a unit quantity of the packing to bind some analyte, often a well characterized protein such as bovine serum albumin. The static loading capacity35 is very different from functional capacity,2 which is the maximum amount of a particular feedstock that can be loaded and still achieve acceptable purification and recovery. Functional capacity is determined empirically for each type of load and associated set... 

Recent data indicate that SR-BI is a nonspecific receptor for many lipophilic molecules (Lorenzi et al., 2008 Reboul et al., 2007b). Apart from HDLs, rodent SR-BI also binds to LDL, VLDL, acetylated LDL, oxidized LDL, and maleylated bovine serum albumin. SR-BII has a similar ligand specificity and function to that of SR-BI (Webb et al., 1998). However, it has been shown that vitamin E (which like carotenoids is carried in the bloodstream mainly by LDL and HDL) is transported more efficiently into the endothelial cells from HDLs than from LDLs (Balazs et al., 2004 Kaempf-Rotzoll et al., 2003 Mardones and Rigotti, 2004). This is in striking contrast to cholesterol, which is taken up much more efficiently from LDLs than HDLs by the RPE to the retina (Tserentsoodol et al., 2006b). It remains to be shown which lipoproteins are the main carriers for carotenoids transported from blood into the RPE. 

Gelamo and Tabak (2000) reported a dramatic decrease in the binding of SDS to reduced bovine serum albumin at pH 9.0 compared to pH 7.0. However, this phenomenon is expected only if disulfide bonds are reduced there is little difference in the charge of the protein at these two pHs if the disulfide bonds are intact. 

Daniel E, Weber G (1966) Cooperative effects in binding by bovine serum albumin. I. The binding of l-anilino-8-naphthalenesulfonate. Fluorimetric titrations. Biochemistry 5 1893-1900... 

Sawada and coworkers [25-27] studied the iso-pH 7.4 MDCK permeabilities of very lipophilic molecules, including chlorpromazine (CPZ) [25], These authors included 3% wt/vol bovine serum albumin (BSA) on the apical (donor) side, and 0.1-3% BSA on the basolateral (acceptor) side, and found that plasma protein binding greatly affected the ability of molecules to permeate cellular barriers. They observed cell tissue retention of CPZ ranging from 65 to 85%, depending on the... 

Figure 6 Effect of the increased rotational correlation time on the proton relaxivity of MP2269, a Gd111 chelate capable of noncovalent protein binding (Scheme 2). The lower NMRD curve was measured in water, whereas the upper curve was obtained in a 10%w/v bovine serum albumin solution in which the chelate is completely bound to the protein. The rotational correlation times calculated are rR=105ps in the nonbound state, and rR= 1,000 ps in the protein-bound state (t=35°C). For this chelate, the water exchange...

Mir MM, Fazili KM, Abul Qasim M. Chemical modification of buried lysine residues of bovine serum albumin and its influence on protein conformation and bilirubin binding. Biochim. Biophys. Acta 1992 1119 261-267. 

Purify the iodinated protein from excess reactants by gel filtration using a desalting resin. The column may be pre-treated by passing a solution of bovine serum albumin (BSA) through it to eliminate nonspecific binding sites that could cause significant protein loss in small-sample applications. 

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.

Sahyun, M.R.V. (1966) Binding of aromatic compounds to bovine serum albumin. Nature 209, 613-614. 

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... 

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