University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Email: schalk@unf.edu
Website: @unf

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Plasma Mouse

Classification: Biological fluid -> blood -> plasma -> mouse

Citations 3

"Monitoring Of Mouse Immunoglobulin G By Flow Injection Analytical Affinity Chromatography"
Anal. Chim. Acta 1991 Volume 245, Issue 1 Pages 1-6
W. Stöcklein, V. Jäger and R. D. Schmid

Abstract: Oxiran acrylic beads (Sigma) (10 mg) were incubated at room temperature for 15 h with 1 M phosphate buffer (pH 7.5) (200 µL) and antibody (rabbit, sheep or goat anti-mouse IgG) or protein A solution (40 to 300 µL), corresponding to 0.5 mg of protein. Remaining oxiranyl groups were blocked by addition of 1 M ethanolamine (1 ml) and further incubation (2 h). The analytical column (1.5 cm x 1 mm) was filled with the prepared beads and formed part of the flow injection system. The procedure comprised: injection of sample or standard (35 µL), diluted as required with phosphate-buffered saline containing 0.1% of bovine serum albumim, 20 mM NaN3 and 0.1% of thiomersal; washing with phosphate-buffered saline containing 20 mM NaN3 and 0.1% of 6-aminohexanoic acid; and elution with 0.1 M Na citrate (pH 3.0 for protein A, pH 2.5 for antibodies) containing 20 mM NaN3. Detection was by fluorescence at 280 nm (excitation) and 360 nm (emission). The calibration graph was rectilinear up to 200 µg mL-1 of mouse IgG standard with use of protein A and up to 75 µg mL-1 with use of antibodies. Prerequisites for accurate determination are: the use of protein A as ligand if the mouse IgG sub-class is 2a, b or 3 and if calibration graphs already exist for these sub-classes; the use of an IgG standard from the same hybridoma line, irrespective of the ligand; or the use of a well-characterized mixture of anti-mouse IgG antibodies with equal affinities for each sub-class.
Immunoglobulin G LC Fluorescence Buffer

"Flow Injection Renewable Surface Immunoassay: A New Approach To Immunoanalysis With Fluorescence Detection"
Anal. Chem. 1994 Volume 66, Issue 11 Pages 1825-1831
Cy H. Pollema and Jaromir Ruzicka

Abstract: Agarose beads (~35 µm) coated with goat anti-mouse IgG1 (heavy chain specific) were diluted 1:20 in 0.01 M phosphate buffer containing 0.5 M NaCl (buffer A) to give a suspension containing ~2 x 105 beads/ml. Portions (42 µL) of the suspension were pumped into the jet ring cell of the sequential injection system (diagram given). The beads were retained on a optical flat surface monitored by fluorescence microscopy with excitation at 450-490 nm and use of a 520 nm long-pass emission filter. For competitive assays, a mixture of unlabelled (sample) and R-Phycoerythrin-conjugated mouse IgG1 mAb diluted in buffer A was then pumped into the cell. Unbound sample was washed away, the signal change due to the bound labelled sample was measured, and the beads were then removed from the cell with a reversed buffer flow; buffer A was used as the carrier throughout (0.25-1 ml/min). The calibration graph was linear from 1-5 µg/ml of mouse IgG1 mAb (using 5 µg/ml of labelled antigen and a contact time of 25 s). The method was also applied to a non-competitive immunoassay (details given).
Immunoglobulin G Fluorescence Immunoassay Agarose beads Sequential injection Immobilized protein Buffer Jet ring cell Renewable surface

"Determination Of Immunostimulative Acylpeptide (FK 565) [N-(1-oxoheptyl)-D-γ-glutamyl-L-erythro-α,e-diaminopimelyl-D-alanine] In Plasma By HPLC With Fluorogenic Post-column Derivatization"
Bunseki Kagaku 1987 Volume 36, Issue 11 Pages 787-791
Tashiro, Y.;Suzuki, A.;Noda, K.;Noguchi, H.

Abstract: Rat or mouse plasma was deproteinized with 6% trichloroacetic acid solution and the supernatant solution was applied to a Bond-Elut C18 cartridge. The retained FK 565 was eluted with mobile phase [20 mM phosphate buffer (pH 2.5) - acetonitrile (79:21)] at 0.5 mL min-1 and was determined on a TSK-gel LS-410 ODS Sil column (15 cm x 4 mm) with fluorimetric detection by using post-column derivatization with phthalaldehyde in the presence of 2-mercaptoethanol. The coefficient of variation were <2.3%. The limits of determination were 50 and 20 ng mL-1 of FK 565 in rat and mouse plasma, respectively.
HPLC Fluorescence Post-column derivatization