scholarly journals Surface isoelectric focusing (sIEF) with carrier ampholyte pH gradient

2017 ◽  
Vol 38 (20) ◽  
pp. 2565-2575 ◽  
Author(s):  
Zhichao Wang ◽  
Cornelius Ivory ◽  
Adrienne R. Minerick
Keyword(s):  
Isoelectric Focusing ◽  
Ph Gradient ◽  
Carrier Ampholyte

scholarly journals Immobilized pH gradient focusing of alkaline proteins: analysis of the isoform composition of purified human non-secretory ribonucleases from kidney, liver and spleen

1993 ◽  
Vol 296 (3) ◽  
pp. 553-556 ◽  
Author(s):  
E C Coronel ◽  
B W Little ◽  
J A Alhadeff
Keyword(s):  
Western Blotting ◽  
Isoelectric Focusing ◽  
Polyclonal Antibodies ◽  
Ph Gradient ◽  
Carrier Ampholyte ◽  
Marker Proteins ◽  
Methodological Problems ◽  
Kidney Liver ◽  
Suitable Technique ◽  
Basic Region

Previous studies on the isoform composition of human ribonucleases (RNAases) have resulted in confusing and inconsistent results, presumably due to methodological problems in electrofocusing of alkaline proteins. In the present study, immobilized pH gradient (IPG) carrier ampholyte (CA) isoelectric focusing (IEF) and conventional CA-IEF have been evaluated for the analysis of the isoforms of human non-secretory RNAases purified from kidney, liver and spleen. CA-IEF proved unsuitable since the alkaline RNAase isoforms migrated into the cathode. IPG-CA-IEF, however, resolved the RNAase isoforms and marker proteins in the basic region of the gel matrix. The three RNAases had comparable isoform profiles, each with two protein bands with approximate pI values of 10.3 and 10.4. Western blotting showed that the two protein bands of each RNAase were immunoreactive (with polyclonal antibodies that recognize RNAase), indicating that the protein bands are RNAase isoforms. The present results provide reliable pI data on human RNAase isoforms and suggest that IPG-CA-IEF should be a suitable technique for analysing the isoforms of other alkaline proteins.

Instability and non-linearity of the pH gradient formed in isoelectric focusing

1979 ◽  
Vol 174 (1) ◽  
pp. 1-11 ◽  
Author(s):  
A. Murel ◽  
I. Kirjanen ◽  
O. Kirret
Keyword(s):  
Isoelectric Focusing ◽  
Ph Gradient

Focusing of Proteins in a Horseshoe Microchannel

2008 ◽  
Author(s):  
Jaesool Shim ◽  
Prashanta Dutta ◽  
Cornelius F. Ivory
Keyword(s):  
Capillary Electrophoresis ◽  
Electric Field ◽  
Isoelectric Focusing ◽  
Ph Gradient ◽  
Complex Geometries ◽  
Electrokinetic Phenomena ◽  
Double Peaks ◽  
Zone Electrophoresis ◽  
Carrier Ampholytes ◽  
Ph Range

Ampholyte based isoelectric focusing (IEF) simulation was conducted to study dispersion of proteins in a horseshoe microchannel. Four model proteins (pls = 6.49, 7.1, 7.93 and 8.6) are focused in a 1 cm long horseshoe channel under an electric field of 300 V/cm. The pH gradient is formed in the presence of 25 biprotic carrier ampholytes (ΔpK = 3.0) within a pH range of 6 to 9. The proteins are focused at 380 sec in a nominal electric field of 300 V/cm. Our numerical results show that the band dispersions of a protein are large during the marching stage, but the dispersions are significantly reduced when the double peaks start to merge. This rearrangement of spreading band is very unique compared to linear electrokinetic phenomena (capillary electrophoresis, zone electrophoresis or electroosmosis) and is independent of channel position and channel shape. Hence, one can perform IEF in complex geometries without incorporating hyperturns.

Isoelectric Focusing in Infinite-Component Mixtures. Creation of a pH Gradient

1989 ◽  
pp. 181-210
Author(s):  
V. G. Babskii ◽  
M. Yu. Zhukov ◽  
V. I. Yudovich
Keyword(s):  
Isoelectric Focusing ◽  
Ph Gradient ◽  
Infinite Component
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