ampholyte


Also found in: Dictionary, Medical.

ampholyte

[′am·fə‚līt]
(chemistry)
An amphoteric electrolyte.
References in periodicals archive ?
Secretome samples (80 [micro]g proteins) were mixed with a rehydration buffer (7 M urea, 2M thiourea, 2% CHAPS, 120mM DTT, 2% ampholyte pH 3-10, and bromophenol blue) and rehydrated into a 7 cm nonlinear immobilized pH gradient (IPG) strips of pH range 3-10 (GE healthcare) for 16-18 hours at room temperature.
Each ReadyStrip[TM] IPG strip (pH 3-10 nonlinear, 17 cm) (Bio-Rad, USA) was passively rehydrated overnight at room temperature with 120 pig of protein sample, which was earlier obtained through appropriate dilution with rehydration buffer (7 M urea, 2 M thiourea, 4% w/v CHAPS, 0.5% v/v of Biolyte 3/10 Ampholyte 40%, 20 mM DTT and 0.002% w/v bromophenol blue).
It was established that synthesized ampholytes possess sufficient high sorption and complexing ability in relation to ions of copper, nickel, uranyl both from sour and alkaline solutions.
Two-dimensional protein electrophoresis was undertaken using constant protein loading as previously described (5,6) using an ampholyte pH range 3.5 to 10.0.
For 2-dimensional electrophoresis, 500 ([micro]g of plasma proteins were diluted in a buffer containing 8 mol x [L.sup.-1] urea, 2% CHAPS (wt/vol), 40 mol x [L.sup.-1] dithiothreitol, 0.2% Bio-Lyte ampholyte (Bio-Rad Labs, Hercules, CA), and 0,01 % (wt/vol) bromophenol blue.
The protein pellet was suspended in appropriate volume of 2D rehydration buffer [8M urea, 2% CHAPS (wt/vol), 50 mM dithiothreitol (DTT), 0.2% Bio-Lyte 3/10 ampholyte, 0.001% bromophenol blue] (BIO-RAD, Hercules, CA, USA) and the protein concentration was estimated by Bradford method (13) using bovine serum albumin as standard.
The stock solution of acrylamide was prepared by mixing 30% acrylamide powder with 5.2% bis acrylamide, 10% NP-40, 10% CHAPS, 10% APS, 20mM NaOH as catholyte, 0.01[micro]M ortho phosphoric acid as anolyte, 8M urea and 250[micro]l of ampholyte as sample overlay.
Initial phenotype ascertainment for any hemoglobin with anomalous electrophoretic migration besides Hb S, was done by isoelectrofocusing in 5.05% polyacrylamide gel with ampholyte pH 7-9, according to standard procedures (8).
The pellet was re-suspended in 300 pl of 2D sample rehydration solution containing 7 M urea (Sigma, USA), 2M ThioUrea (Sigma, USA), 0.2% pH 3-10 linear IPG Ampholyte (Bio-Rad Laboratories, USA), 4% CHAPS (Sigma, USA), 1% HED (2-hydroxyethyldisulfide, Sigma, USA), and 1% DTT (Dithiothreitol, Sigma, USA).
Two dimensional electrophoresis (2-DE): Four lenses were homogenized in lysis buffer (0.5 M Tris-HCl pH 6.8, 0.25 M EDTA, Urea, 0.5 M dithiothreitol (DTT), Glycerol, NP40, ampholyte buffer pH 3-10) and centrifuged at 15,000 x g for 20 min at 4C.
The strips were rehydrated with 137.5 [micro]L of a solution containing 150 [micro]g of protein, 40 mM DTT, 2% ampholyte (IPG Buffer pH3-10) and Destreak rehydration solution (GE Healthcare).
Briefly, cells were dissolved in lysis buffer (7M urea, 2M thiourea, 4% CHAPS, 100 mM DTT, 0.2% pH 3-10 ampholyte, Bio-Rad, USA) in presence of protease inhibitor (Sigma).