Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250

A simplified methodology for the quantitative electroelution of proteins from polyacrylamide gels is described. After staining with Coomassie Brilliant Blue R 250, the identified bands are excised from the gel and the proteins eluted using a procedure developed for use in conventional tube gel electrophoresis equipment.

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Isoelectric focusing of gamma globulins in cerebrospinal fluid from patients with multiple sclerosis.

Clinical Chemistry ◽

10.1093/clinchem/23.12.2213 ◽

1977 ◽

Vol 23 (12) ◽

pp. 2213-2215 ◽

Cited By ~ 16

Author(s):

J L Trotter ◽

G Banks ◽

P Wang

Keyword(s):

Cerebrospinal Fluid ◽

Isoelectric Focusing ◽

Clinical Laboratory ◽

Clinical Analysis ◽

Globulin Fraction ◽

Ammonium Sulfate Precipitation ◽

Constant Concentration ◽

Brilliant Blue G ◽

Coomassie Brilliant Blue ◽

Coomassie Brilliant

Abstract The technique of isoelectric focusing has been adapted for rapid clinical analysis for globulins in cerebrospinal fluid with use of commercially prepared horizontal-slab acrylamide gels. The globulin fraction is concentrated by ammonium sulfate precipitation, which allows more of the relevant protein to be applied, use of a wider range of total protein concentrations, and higher resolution than is true for previously described methods. Critical variables include a constant concentration and volume of IgG, a constant low temperature of the acrylamide gel, and sensitive staining with Coomassie Brilliant Blue G-250. The apparatus used is adaptable for other electrophoretic procedures in the clinical laboratory, and the use of commercially prepared gel slabs is more convenient, more reproducible, and requires less time than other methods.

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MEASUREMENT OF CANINE PITUITARY PROLACTIN

Journal of Endocrinology ◽

10.1677/joe.0.0560245 ◽

1973 ◽

Vol 56 (2) ◽

pp. 245-NP ◽

Cited By ~ 5

Author(s):

P. G. SALUJA ◽

M. GRONOW ◽

J. M. HAMILTON

Keyword(s):

Isoelectric Focusing ◽

Isoelectric Point ◽

Gel Electrophoresis ◽

Polyacrylamide Gel ◽

Brilliant Blue ◽

Coomassie Brilliant Blue ◽

Ovine Prolactin ◽

Pituitary Prolactin ◽

Coomassie Brilliant

SUMMARY Isoelectric focusing in polyacrylamide gel followed by staining with Coomassie brilliant blue was used for the densitometric estimation of ovine prolactin standard and canine pituitary prolactin. The results were compared with those obtained by conventional polyacrylamide gel electrophoretic procedures and isoelectric focusing was found to be a valid technique for the estimation of prolactin and to offer greater technical convenience. The mobility of ovine and canine prolactin was similar in isoelectric focusing and gel electrophoresis. The isoelectric point of ovine and canine prolactin was found to be 6·17 and 6·61 respectively. Constant levels of prolactin were found in the pituitaries of bitches at oestrus.

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