Recombinant fluorescence detection

Two different types of pulsed EPR experiments are possible a spectrum can be measured at a fixed time after the pulse by variation of the field strength B (Eq. 72), or the time profile of a particular spectral line can be measured at constant B to give kinetic information. One vziriation of this kinetic method is to detect the recombination of singlet-state radical ion pairs in liquid hydrocarbons by the fluorescence of the product excited state [142]. This technique is known as fluorescence-detected magnetic resonance (FDMR) and provides information on the spin dynamics of the radical ion pair as well as the chemical kinetics. 

It was shown that the chromatography method was suitable to separate recombinant EPO from amounts of human serum albumin commonly present as a stabilizer in various pharmaceutical preparations. In addition, it was possible to obtain different elution profiles for EPO products with variations in the glycoforms. Fluorescence detection was applied for quantification and showed linear signals over the range of 10-200-pg/mL EPO. 

Kamoda, S., Ishikawa, R., and Kakehi, K., Capillary electrophoresis with laser-induced fluorescence detection for detailed studies on N-linked oligosaccharide profile of therapeutic recombinant monoclonal antibodies, J. Chromatogr. A, 1133, 332-339, 2006. 

Ma, S. and Nashabeh, W. Carbohydrate analysis of a chimeric recombinant monoclonal antibody by capillary electrophoresis with laser-induced fluorescence detection. Anal Chem, 71, 5185,1999. 

The ionization energy of TMPD in solid and liquid solutions of hydrocarbons was studied by biphotonic ionization and detection of recombination fluorescence (Bullot and Gauthier, 1976). Generally, the photoionization threshold decreased on going from the solid to the liquid phase. The following sequence was observed ... 

In addition to measuring total recombination coefficients, experimentalists seek to determine absolute or relative yields of specific recombination products by emission spectroscopy, laser induced fluorescence, and optical absorption. In most such measurements, the products suffer many collisions between their creation and detection and nothing can be deduced about their initial translational energies. Limited, but important, information on the kinetic energies of the nascent products can be obtained by examination of the widths of emitted spectral lines and by... 

Many of the methods developed to study protein interactions use the bait/prey model to detect interacting partners (Phizicky and Fields, 1995 Archakov et al., 2003 Piehler, 2005). The bait protein is a purified protein (often recombinant) that is used to lure and capture a putative interacting protein or biomolecule. The bait protein may be immobilized to a solid phase for affinity separations or be used in solution. It also may be fusion tagged (i.e., GST or 6X His) or labeled with a detectable molecule, such as a fluorescent probe. It often is the case... 

Fig. 11.2 Localization of GFP and calsequestrin (CSQ) in neonatal rat cardiac myocytes. Myo cytes were infected with a recombinant adenovirus containing the cDNAs of GFP and CSQ. Both cDNAs were expressed under control of a separate cytomegalovirus promoter. Expression of CSQ and the coexpressed GFP was detected by fluorescence microscopy. Left-. GFP fluorescence (green), middle immunostaining of CSQ (red), right overlay of GFP fluorescence and immunos taining. Nuclei were counterstained with DAPI (blue). Courtesy of Ulrich Gergs...

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