BSA protein

Figure 8. Comparison of the response of the SPR and MOSPR sensor to the adsorption of BSA proteins (10 pg/ml in PBS).

Fig. 23 Normalized absorption spectra of the free BSA protein (1), BSA-dye 50 conjugate (2) and steady state fluorescence emission spectrum of the BSA-dye conjugate (2 )...

Proteins have also been used as CMPAs for chiral resolution by liquid chromatography. Bovine serum albumin (BSA) and a]-acid glycoproteins (AGP) were investigated as CMPAs. Allenmark et al. [87] used BSA protein as the CMPA for... 

Bovine adrenal capillary endothelial cells Selective wetting of topographical features with BSA, protein backfill 2000 [171]... 

Figure 4.26 Rejection coefficient of bovine serum albumin (BSA) protein as a function of time for untreated titania membrane and two titania membranes treated with 0.1 M phosphoric acid at room temperature for 1 and 14 hours, respectively. (A) without phosphate buffer (B) with 0.01 M Na2HP04/NaH2P04 buffer [Randon el al.. 1995]...

Alternative protocols are described elsewhere (Ribeiro-Neto efal., 1985 Kopf and Woolkalis, 1991 Carty, 1994). ATP, phospholipids and small amounts of certain ionic and nonionic detergents (i.e. SDS, CHAPS, Lubrol PX) promote dissociation of the A and B protomers (Moss et a/., 1986). We do not use SDS since it denatures solubilized G proteins very easily. DTT or 3-ME are necessary to break the disulfide bonds of the A protomer. Supplementation with BSA (final concentration approx. 0.9 mg/ml) helps prevent loss of enzyme through adsorption to the walls of the tube, and ensures recovery of proteins following precipitation with sodium chloride/acetone, trichloroacetic acid (TCA), or chloroform/methanol. Furthermore, when samples are subjected to SDS-PAGE, intensities of the stained 67 kDa BSA protein bands allow rough estimation of incomplete recovery of the precipitated sample (see section 4.5). Preactivated PT should be used immediately, and enzyme left over from an experiment should be discarded, since reduced toxin has been shown to lose activity rapidly (Kaslow et a/., 1989). 

To compare activity between samples, all are normalized for total cytosolic protein. We carry this out using the BioRad protein assay (a commercial preparation of the Bradford assay), and a standard curve from BSA protein standard... 

Bakerbond Application Note Bi-001 Extraction and Concentration of BSA Protein from Dilute aqueous Solution (See Suggested Reading, Chapter 1). 

To investigate the mechanism of the BSA on the improvement of enzymatic hydrolysis of pretreated CWR, the adsorptions of BSA on various substrates were studied [17]. Three sets of 20-ml vials were used to test the adsorption of BSA to Avicel, pretreated CWR, and lignaceous residue, respectively. The mixtures of citrate buffer (pH=4.8) with 8% (w/w) Avicel, 8% (w/w) pretreated CWR, or 3% (w/w) lignaceous residue were preheated to 50°C for 30 min. BSA was then added to the vials with the ratio of BSA to dry solid equal to 0.1 g BSA/g dry solid. The final working volume was 10 ml. One-milliliter aliquots were periodically withdrawn at start and after 1, 2, 4, 8, 24, 48, and 72 h. The aliquots were pretreated according to the procedures described in Effect of Tween 20 and BSA on Enzyme Protein Concentration and Activity for BSA protein concentration measurement. 

The protein concentration in solution was measured by the Bradford protein assay using BSA as a standard (Bio-Rad). The Bradford colorimetric method cannot distinguish the BSA protein from enzyme protein. Therefore, no differentiation was possible between BSA and enzymes in solution other than by measuring the enzyme activity. The enzyme activities in solution including FPU and CBU were measured according to the methods described by Ghose [35]. 

Liu, G., X. Miao, W. Fan, R. Crawford, and Y. Xiao. Porous PLGA microspheres effectively loaded with BSA protein by electrospraying combined with phase separation in liquid nitrogen. Journal of... 

Fig. 12.2 5x5 pm AFM height image of BSA protein adsorbed on HOPG with nanobubbles formed prior to protein adsorption left, image scale 5x5 pm, height scale 10 tun) and BSA removal (surface fraction on axial axis) by electrochemically generated nanobubbles right) (from Z.H. Wu et al. [7])... 

Figure 3.14 Fluorescence emission of frans-4-dimethylamino-4 -amino-stilbene in a free state in 8 x 10 M toluene solution (closed squares) and an immobilized state cross-linked with cyanuric chloride to the coating BSA protein in toluene (open squares) and in glycerin (rhombus) [55], (Reproduced with permission from Elsevier.)...

Fig. 1 Various functional coatings onto porous silicon surfaces (a) thermally responsive polymer/porous silieon hybrid for biosensing (Perelman et al. 2010), (b) porous sihcon/polymer nanoeomposite for biosensing (Li et al. 2005), (c) BSA protein-adsorbed porous silicon surface (Tay et al. 2004), (d) diamond-eapped porous silicon film for optical devices (Fernandes et al. 1999), (e) biocompatible polymer/porous silieon eomposite fiber (Kashanian et al. 2010), (f) block copolymer-coated surface for templating (Qiao et al. 2007), (g) ZnO-deposited porous silicon (Kayahan 2010), (h) SERS active silver-coated porous silicon (Virgaetal. 2012) (Reproduced, with permission, from Perelman et al. (2010), Li et al. (2005), Tay et al. (2004), Fernandes et al. (1999), Kashanian et al. (2010), Qiao et al. (2007), Kayahan (2010), Virga et al. (2012))...

Fig, 3.14. (a) 2D BN-PAGE of bovine serum stained with colloidal Coomassie. (c) S, Cu and Zn bonded to the protein were detected by (b) LA-ICP-MS In five line scans from the (BSA) protein band. 

Approach 3 Functional designer materials with built in element of free radical damage using BSA protein... 

Since, protein adsorption is the first and foremost step when a biomaterial comes in contact with serum, a number of studies have been performed to modify CS for favorable protein adsorption and subsequent cellular behavior in vitro. Chung et al. photochemically grafted adhesive peptides onto the surface of CS and found that the protein-adsorbed CS promotes the adhesion and proliferation of human endothelial cells [76], Recently, bovine serum albumin (BSA) protein was adsorbed onto the surface of CS functionalized with different nanostructured carbon (graphene oxide (GO) and carbon nanohoms). The adsorption of BSA on CS was found to favorably modulate the response of osteoblasts cells, as seen by the attachment and spreading of cells to form an ECM on the protein adsorbed substrate (Fig. 2.7) [77]. 

Monoamine oxidase amperometric biosensor based on SPE were also modified with MWCNT by using the drop casting technique for the determination of antidepressants in model solutions and dosage forms. The authors used BSA protein which provided a matrix for the immobilization of the enzyme and protection of the enzyme activity when glutaraldehyde is used as a linker. Serafin et aZ. developed a label free dual immunosensor for the determination of human growth and prolactin hormones. The electrochemical immunosensor was based on CNT modify carbon SPE platform with the presence of poly(ethylene-dioxythiophene) (PEDOT) and gold nanoparticles. Again, the hybrid nano-material composite facilitated a proper immobilization of the antibody on the electrode matrix. 

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