Microchip electrophoresis utilizing an in situ photopolymerized Phos-tag binding polyacrylamide gel for specific entrapment and analysis of phosphorylated compounds

The Analyst ◽  
2017 ◽  
Vol 142 (18) ◽  
pp. 3416-3423 ◽  
Author(s):  
Sachio Yamamoto ◽  
Miyuki Himeno ◽  
Masaya Kobayashi ◽  
Miki Akamatsu ◽  
Ryosuke Satoh ◽  
...  
Keyword(s):  
Microchip Electrophoresis ◽  
Polyacrylamide Gel ◽  
Phosphorylated Compounds ◽  
Crossing Point

A method was developed for the specific entrapment and separation of phosphorylated compounds using a Phos-tag polyacrylamide gel fabricated at the channel crossing point of a microfluidic electrophoresis chip.

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

Gels ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 268
Author(s):  
Sachio Yamamoto ◽  
Shoko Yano ◽  
Mitsuhiro Kinoshita ◽  
Shigeo Suzuki
Keyword(s):  
Microfluidic Chip ◽  
Tetraacetic Acid ◽  
Improved Method ◽  
Phosphate Ions ◽  
Phosphorylated Compounds ◽  
Phosphorylated Peptides ◽  
Crossing Point ◽  
Acrylamide Solution ◽  
Separation Buffer

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.

Microchip electrophoresis of oligosaccharides using lectin-immobilized preconcentrator gels fabricated by in situ photopolymerization

The Analyst ◽  
2012 ◽  
Vol 137 (9) ◽  
pp. 2211 ◽  
Author(s):  
Sachio Yamamoto ◽  
Sho Suzuki ◽  
Shigeo Suzuki
Keyword(s):  
Microchip Electrophoresis

On-line microchip electrophoresis-mediated preconcentration of cationic compounds utilizing cationic polyacrylamide gels fabricated by in situ photopolymerization

The Analyst ◽  
2018 ◽  
Vol 143 (18) ◽  
pp. 4429-4435 ◽  
Author(s):  
Sachio Yamamoto ◽  
Fuka Okada ◽  
Mitsuhiro Kinoshita ◽  
Shigeo Suzuki
Keyword(s):  
Methyl Methacrylate ◽  
Efficient Method ◽  
Microchip Electrophoresis ◽  
Polyacrylamide Gels ◽  
Poly Methyl Methacrylate ◽  
Cationic Polyacrylamide ◽  
Cationic Compounds ◽  
On Line

A simple and efficient method was developed for the fabrication of a cationic sample preconcentrator on a channel of a commercial poly(methyl methacrylate) (PMMA) microchip.

scholarly journals Isolation of mouse myocardial gap junctions.

1980 ◽  
Vol 86 (3) ◽  
pp. 755-764 ◽  
Author(s):  
R W Kensler ◽  
D A Goodenough
Keyword(s):  
Gap Junctions ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
New Method ◽  
Diffuse Band ◽  
Intercalated Disk

A new method is presented for the isolation of an enriched fraction of mouse myocardial gap junctions without the use of exogenous proteases. The junctions appear well preserved morphologically and similar to their appearance in situ. Contaminants of the preparation include fragments of the fascia adherens region of the intercalated disk. SDS polyacrylamide gel electrophoresis of the preparation reveals seven major bands with apparent mol wt of 28,000; 31,000; 33,500; 43,000; 47,000; 49,000; and 57,000. Only the bands at 38,000; 31,000; 33,500; and possibly the diffuse band at 47,000 copurify with the morphologically assayed gap junctions. Evidence is presented that the peptides at 43,000 and 57,000 are contained within the contaminating fascia adherens.

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