Detection of Low Molecular Weight IReduced Zein Polypeptides Separated by Polyacrylamide Gel Electrophoresis

1986 ◽  
Vol 51 (5) ◽  
pp. 1366-1367
Author(s):  
SHELA GORINSTEIN ◽  
GEORGE V. QUICKE ◽  
ANNE M. PHILLIPS
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Low Molecular Weight

Fractionation of Chromatin Components

1973 ◽  
Vol 51 (5) ◽  
pp. 709-720 ◽  
Author(s):  
John J. Monahan ◽  
Ross H. Hall
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Urea Solution ◽  
Equilibrium Density ◽  
Tissue Culture Cell ◽  
Low Molecular Weight ◽  
Nonhistone Proteins

A general method for isolation and fractionation of chromatin into its four major components, DNA, RNA, histories, and nonhistone proteins, is described. The procedure avoids the use of strongly acidic or alkaline conditions, or the use of ionic detergents or phenol. As few as 14 × 106 cells can be used. The procedure is reasonably rapid and has been used successfully with a number of tissue culture cell lines. The chromatin components are dissociated in a 3 M NaCl – 5 M urea solution containing 2-mercaptoethanol and EDTA. The DNA and high molecular weight RNA are collected by high-speed centrifugation and DNA is separated from the RNA by means of Cs2SO4 equilibrium density centrifugation. The histones, nonhistone proteins, and low molecular weight RNA's are fractionated using DEAE-cellulose column chromatography and polyacrylamide gel electrophoresis. A small amount (< 1%) of protein is present in the DNA and RNA fractions. At least 11 low molecular weight RNA subfractions can be detected by means of polyacrylamide gel electrophoresis.

Method for measuring the concentration of urinary proteins according to their molecular size category.

1976 ◽  
Vol 22 (5) ◽  
pp. 667-672 ◽  
Author(s):  
A J Pesce ◽  
A Hsu ◽  
C Kornhauser ◽  
K Sethi ◽  
B S Ooi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Low Molecular Weight ◽  
Tubular Proteinuria ◽  
Equal Concentration ◽  
Glomerular Proteinuria ◽  
Low Molecular Weight Proteins

Abstract We combined the use of a concentrating device (Minicon) and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate to semi-quantitate the concentration of (a) the collective low-molecular-weight proteins and (b) of albumin excreted in the urine of patients after renal transplantation. Analytical recovery of many serum proteins from samples concentrated 100-fold in the Minicon apparatus was about 70%. It was possible to examine many urine samples by polyacrylamide gel electrophoresis after concentration with this device. The reproducibility (CV) of the technique was on the order of 20% when albumin and low-molecular-weight protein were in about equal concentration. The method was adequate to differntiate glomerular and tubular proteinuria, because in glomerular proteinuria the ratio of albumin to low-molecular-weight proteins is about 20/1, whereas in tubular proteinuria the ratio is about 1/1.

The RNA Fractions of Chromatin from L-Cells

1974 ◽  
Vol 52 (10) ◽  
pp. 922-935 ◽  
Author(s):  
John J. Monahan ◽  
Ross H. Hall
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Weight Fraction ◽  
Polyacrylamide Gel ◽  
Low Molecular Weight ◽  
Two Dimensional ◽  
Cell Nuclei ◽  
L Cell

Chromatin obtained from L-cell nuclei has been shown to contain three distinct RNA fractions. One, a low molecular weight RNA fraction (3–7 S), has been isolated and purified. Two-dimensional polyacrylamide gel electrophoresis indicates that this fraction contains at least 28 diverse RNA components. Both the rate of incorporation of [32P]H3PO4 into this fraction and the loss of [3H]uridine from the fraction after a 1 h pulse suggest that these RNA's are slowly synthesized and are very stable. Not all the species within the low molecular weight fraction have the same turnover rates.A high molecular weight RNA fraction (15–30 S) has also been isolated from L-cell chromatin. From the rate of [32P]H3PO4 incorporation and the loss of [3H]uridine from this fraction after a 1 h pulse, it appears that this RNA fraction is rapidly synthesized and is either unstable or is rapidly removed from chromatin once synthesized.A third RNA fraction also present in L-cell chromatin appears to be tightly bound to the DNA. It has been isolated and shown to be of low molecular weight (3–5 S). This RNA also appears to be rapidly synthesized and to be metabolically unstable.

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