A simple method of labelling mouse Thy-1 antibodies with FITC

Gani, M.; Hunt, Timothy J.; Summerell, J

doi:10.1016/0022-1759(80)90167-2

Cited by 13 publications

(6 citation statements)

References 14 publications

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“…We applied this method to analyses of 8-aminopyrene-1,3,6-trisulfonate derivatives of oligosaccharides; in addition, FITC-labeled glycopeptides and glycoproteins were prepared using the standard method. 50 As demonstrated in Figure 6, a careful choice of voltage setting and direction of sample concentration and separation enables sensitive detection with high efficiency and without loss of resolution efficiency. Herein, we described only the concentration of anionic samples.…”

Section: Discussionmentioning

confidence: 99%

In Situ Fabrication of Ionic Polyacrylamide-Based Preconcentrator on a Simple Poly(methyl methacrylate) Microfluidic Chip for Capillary Electrophoresis of Anionic Compounds

2008

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A simple and efficient method was developed for fabrication of an anionic sample preconcentrator on a channel of a commercial poly(methyl methacrylate) (PMMA)-made microchip using no photolithography or etching technique. The originality of our preconcentrator is based on simple photochemical copolymerization of monomers using the following procedure: All channels of the PMMA-made microchip were filled with gel solution comprising acrylamide, N,N'-methylene-bisacrylamide, and 2-acrylamide-2-methylpropanesulfonic acid with riboflavin as a photocatalytic initiator. In situ polymerization near the cross of the sample outlet channel was performed by irradiation with an argon ion laser beam, which is also used as the light source for fluorometric detection. The electrokinetic property and electric repulsion between sample components and anionic groups on the polyacrylamide gel layer produce, trap, and concentrate anions within a few minutes at the interface of the cathodic side of the gel layer. This method displays concentration factors as high as 10 (5). The availability of ionic preconcentrator was demonstrated by applying sensitive analysis of oligosaccharides labeled with 8-aminopyrene-1,3,6-trisulfonate and some glycoproteins labeled with fluorescein isothiocyanate under various buffer systems.

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Section: Discussionmentioning

confidence: 99%

In Situ Fabrication of Ionic Polyacrylamide-Based Preconcentrator on a Simple Poly(methyl methacrylate) Microfluidic Chip for Capillary Electrophoresis of Anionic Compounds

2008

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“…The monophosphorylated β-casein was derivatized with FITC according to a previously method [ 35 ] with slight modifications. A solution of 5 μL of 20 mM FITC in acetone was mixed with 1 mM β-casein phosphorylated peptides in 90 μL of 10 mM sodium phosphate (pH 9.4).…”

Section: Methodsmentioning

confidence: 99%

In Situ Pinpoint Photopolymerization of Phos-Tag Polyacrylamide Gel in Poly(dimethylsiloxane)/Glass Microchip for Specific Entrapment, Derivatization, and Separation of Phosphorylated Compounds

Yamamoto

Yano

Kinoshita

et al. 2021

Gels

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An improved method for the online preconcentration, derivatization, and separation of phosphorylated compounds was developed based on the affinity of a Phos-tag acrylamide gel formed at the intersection of a polydimethylsiloxane/glass multichannel microfluidic chip toward these compounds. The acrylamide solution comprised Phos-tag acrylamide, acrylamide, and N,N-methylene-bis-acrylamide, while 2,2′-azobis[2-methyl-N-(2-hydroxyethyl)propionamide] was used as a photocatalytic initiator. The Phos-tag acrylamide gel was formed around the channel crossing point via irradiation with a 365 nm LED laser. The phosphorylated peptides were specifically concentrated in the Phos-tag acrylamide gel by applying a voltage across the gel plug. After entrapment of the phosphorylated compounds in the Phos-tag acrylamide gel, 5-(4,6-dichlorotriazinyl)aminofluorescein (DTAF) was introduced to the gel for online derivatization of the concentrated phosphorylated compounds. The online derivatized DTAF-labeled phosphorylated compounds were eluted by delivering a complex of phosphate ions and ethylenediamine tetraacetic acid as the separation buffer. This method enabled sensitive analysis of the phosphorylated peptides.

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“…Preparation of FITC-Labeled Cytochrome C: The FITC-labeled cytochrome C was prepared according to previously reported protocols. [24,41] A solution of 1 mg of equine heart cytochrome C (12 kDa, Sigma Aldrich, USA) plus 20 mm of sodium phosphate buffer with pH 9.4 (90 µL) (Alfa Aesar, USA) was mixed with 5 µL of 20 mm of FITC (Sigma Aldrich, USA) in acetone (99%, Samchun, South Korea). The solution was heated at 50 °C for 1 h and diluted in 9.9 mL of 0.1 mm of NaCl (Samchun, South Korea) solution for analysis.…”

Section: Methodsmentioning

confidence: 99%

A Multiscale‐Porous Anion Exchange Membrane for Convenient and Scalable Electrokinetic Concentration of Cationic Species

Lee

Kwon

Lim

2020

Adv Funct Materials

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A device able to electrokinetically concentrate cationic samples has many potential medical and industrial applications, but until now has remained undeveloped due to the lack of a commercial anion‐permselective material leading to a prohibitively complex fabrication procedure. Herein, a novel multiscale‐porous anion exchange membrane (MP‐AEM) that enables the convenient and scalable electrokinetic concentration of cationic species is proposed. A mechanically enhanced multiscale‐porous structure with a solid framework is realized by adopting polyester resin as an additive to overcome the intrinsic limitations of the AEM material. The scalable MP‐AEM‐embedded electrokinetic concentrator is devised based on the peculiar properties of the MP‐AEM that for allow both ion and fluid transport. With the MP‐AEM, the concentrator is fabricated in a highly streamlined manner consisting only of a simple insertion and assembly. The concentration performance of the MP‐AEM‐embedded electrokinetic concentrator is demonstrated with a positively charged fluorescent dye and a fluorescein‐labeled protein, and the results show enrichment factors of 250 and 500, respectively. The MP‐AEM makes cationic electrokinetic concentration more accessible and scalable, thereby enabling further progress in a wide range of fields.

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A simple method of labelling mouse Thy-1 antibodies with FITC

Cited by 13 publications

References 14 publications

In Situ Fabrication of Ionic Polyacrylamide-Based Preconcentrator on a Simple Poly(methyl methacrylate) Microfluidic Chip for Capillary Electrophoresis of Anionic Compounds

In Situ Fabrication of Ionic Polyacrylamide-Based Preconcentrator on a Simple Poly(methyl methacrylate) Microfluidic Chip for Capillary Electrophoresis of Anionic Compounds

In Situ Pinpoint Photopolymerization of Phos-Tag Polyacrylamide Gel in Poly(dimethylsiloxane)/Glass Microchip for Specific Entrapment, Derivatization, and Separation of Phosphorylated Compounds

A Multiscale‐Porous Anion Exchange Membrane for Convenient and Scalable Electrokinetic Concentration of Cationic Species

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