Microscale multisample two-dimensional electrophoresis of proteins in human serum, cerebrospinal fluid, and urine.

1982 ◽  
Vol 28 (4) ◽  
pp. 824-827 ◽  
Author(s):  
T Manabe ◽  
E Hayama ◽  
T Okuyama
Keyword(s):  
Cerebrospinal Fluid ◽  
Human Serum ◽  
Distribution Patterns ◽  
Blue Staining ◽  
Two Dimensional ◽  
Protein Distribution ◽  
Two Dimensional Electrophoresis ◽  
Coomassie Blue Staining ◽  
Coomassie Blue ◽  
Dimensional Electrophoresis

Abstract In this technique, in which no denaturing agent is used, proteins in human serum, cerebrospinal fluid, and urine are separated by isoelectric focusing in cylindrical 40 g/L polyacrylamide gels of capillary size (1.3 x 35 mm) for 40 min, followed by electrophoresis in 40--170 g/L polyacrylamide linear gradient gel, with use of 38 x 35 x 1 mm slab gel, for 1 h. Only 2 microL of untreated human serum is required to obtain clear protein-distribution patterns, made visible by Coomassie Blue staining. By use of silver staining, proteins in unconcentrated cerebrospinal fluid can be made visible. An apparatus we devised for microscale two-dimensional electrophoresis enables us to analyze eight protein samples simultaneously.

Computerized quantitative analysis of coomassie-blue-stained serum proteins separated by two-dimensional electrophoresis.

1989 ◽  
Vol 35 (12) ◽  
pp. 2297-2304 ◽  
Author(s):  
A Burgess-Cassler ◽  
J J Johansen ◽  
D A Santek ◽  
J R Ide ◽  
N C Kendrick
Keyword(s):  
Serum Proteins ◽  
Protein Isoforms ◽  
Blue Staining ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Coomassie Blue Staining ◽  
Coomassie Blue ◽  
Single Standard ◽  
Microcomputer Software ◽  
Dimensional Electrophoresis

Abstract Two-dimensional electrophoresis in combination with Coomassie Blue staining was refined for use as a quantitative method. Microcomputer software was developed for use with the IBM AT and compatible computers for analyzing the gels. To test the refined method to determine its usefulness in simultaneous measurements of 28 human serum proteins, we measured each protein relative to a single standard (bovine serum albumin) polymerized at different concentrations in a calibration scale, rather than using 28 individual standards. All samples were analyzed in triplicate. We evaluated calibration, linearity of response, recoveries, units, within-run CV, and between-run CV. The five isoforms of apolipoprotein A-I were analyzed in samples from 16 healthy donors and the isoform ratios determined. The method as presented here should prove useful for diagnosis of non-urgent disease states and for analysis for protein isoforms in relation to disease; it should also be applicable to assays of proteins in other fluids and tissues.

Two-dimensional electrophoresis of cerebrospinal fluid proteins in multiple sclerosis and various neurological diseases

1984 ◽  
Vol 5 (4) ◽  
pp. 236-245 ◽  
Author(s):  
Michael G. Harrington ◽  
Cari R. Merril ◽  
David Goldman ◽  
Xian-Hau Xu ◽  
Dale E. McFarlin
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Neurological Diseases ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Cerebrospinal Fluid Proteins ◽  
Dimensional Electrophoresis

Analytical electrofocusing and two-dimensional electrophoresis of proteins extracted from the mycelia of aggressive and nonaggressive strains of Ophiostoma ulmi

1986 ◽  
Vol 64 (9) ◽  
pp. 2073-2081 ◽  
Author(s):  
Robert S. Jeng
Keyword(s):  
Dutch Elm Disease ◽  
Two Dimensional ◽  
Polyacrylamide Gels ◽  
Ophiostoma Ulmi ◽  
Two Dimensional Electrophoresis ◽  
Coomassie Blue ◽  
Isoelectric Points ◽  
Additional Protein ◽  
Dimensional Electrophoresis ◽  
Electrophoresis Of Proteins

Soluble mycelial proteins from Ophiostoma ulmi (Buism.) Nannf., the causal agent of Dutch elm disease, were separated by analytical electrofocusing and two-dimensional electrophoresis in polyacrylamide gels. Results showed the aggressive and nonaggressive strains of this pathogen each had about 60 Coomassie blue stained bands having isoelectric points from 3 to 7. Both strains of this fungus had their own characteristic electrofocusing patterns. Nonaggressive isolate S116, for example, lacked two protein bands, one near the anode and one near the cathode, but it had five additional protein bands distributed from pH 4 to 6. Two-dimensional electrophoresis of total soluble proteins depicted that there were 36 proteins found to be specific for the nonaggressive isolate S116 and 12 proteins for the aggressive isolate RR2.

Two-Dimensional Electrophoresis of Pseudocholinesterase Components in Normal Human Serum

Nature ◽  
1962 ◽  
Vol 196 (4861) ◽  
pp. 1296-1298 ◽  
Author(s):  
H. HARRIS ◽  
D. A. HOPKINSON ◽  
E. B. ROBSON
Keyword(s):  
Human Serum ◽  
Normal Human Serum ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Normal Human ◽  
Dimensional Electrophoresis

Two-dimensional electrophoresis analysis of human serum proteins during the acute-phase response

1992 ◽  
Vol 13 (1) ◽  
pp. 743-746 ◽  
Author(s):  
Luca Bini ◽  
Barbara Magi ◽  
Carla Cellesi ◽  
Aldo Rossolini ◽  
Vitaliano Pallini
Keyword(s):  
Human Serum ◽  
Acute Phase ◽  
Acute Phase Response ◽  
Serum Proteins ◽  
Phase Response ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis ◽  
Human Serum Proteins

Enhancement techniques for detecting trace and fluid-specific components in two-dimensional electrophoresis patterns.

1982 ◽  
Vol 28 (4) ◽  
pp. 759-765 ◽  
Author(s):  
G B Dermer ◽  
L M Silverman ◽  
J F Chapman
Keyword(s):  
Cerebrospinal Fluid ◽  
Serum Proteins ◽  
Prostatic Acid Phosphatase ◽  
Two Dimensional ◽  
Cibacron Blue ◽  
Two Dimensional Electrophoresis ◽  
Prostatic Fluid ◽  
Creatine Kinase B ◽  
Malignant Effusions ◽  
Dimensional Electrophoresis

Abstract Albumin and other serum-derived proteins were removed from several types of body fluids by affinity chromatography, to facilitate detection of trace or non-serum-derived proteins in two-dimensional electrophoresis patterns. Albumin was removed by the dye Cibacron Blue F3G-A coupled to Sepharose. Two-dimensional patterns of albumin-depleted serum lack the large albumin spot, and several families of spots become visible that ordinarily are partly or totally hidden by it. However, other proteins also bind to Cibacron Blue. Most serum proteins, including albumin, were effectively removed by anti-human serum antibodies coupled to Sepharose. Two-dimensional patterns of serum-depleted cerebrospinal fluid exhibit five clusters of probable nervous-system protein families not detected in serum. One additional family, probably antigenically related to transferrin, was removed by the affinity step. Two-dimensional patterns of serum-depleted prostatic fluid exhibit five major non-serum families, two of which may be creatine kinase B subunits and prostatic acid phosphatase. Two-dimensional patterns of serum-depleted malignant effusions exhibit one or more of three proteins that possibly are tumor products. Pattern matching suggests the presence of one non-serum-derived protein family common to cerebrospinal fluid, prostatic fluid, and malignant effusions. Prostatic fluid and malignant effusions have in common as many as three non-serum families of proteins.

Sign in / Sign up

Export Citation Format

Share Document