EurekaMag.com logo
+ Site Statistics
References:
53,214,146
Abstracts:
29,074,682
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Preparation of protease free and rnase free pancreatic dnase ec 3.1.4.5


Journal of Biological Chemistry 253(20): 7216-7219
Preparation of protease free and rnase free pancreatic dnase ec 3.1.4.5
When pancreatic DNase I[EC 3.1.4.5] is used as a specific biochemical reagent in the preparation of nuclear RNA or nuclear proteins, freedom from contaminating RNases or proteases is an important property of the enzyme preparation. A simple 1-step procedure was developed to effect complete removal of trypsin, chymotrypsin and chymotrypsinogen by a combination of affinity chromatography and salting-out adsorption on lima bean protease inhibitor coupled to Sepharose (a column (0.9 .times. 60 cm) operated in series with a regeneratable 1 ml bed). Commercial preparations of DNase (about 10 mg) give a quantitative yield of the enzyme that is protease-free as evidenced by full stability for more than 10 days at pH 8 and 37.degree. C. Removal of the last traces of RNase was accomplished by affinity chromatography on a column (0.4 .times. 72 cm) of 5'-(4-aminophenyl-phosphoryl)-uridine 2'(3')-phosphate-Sepharose; the product is a highly active DNase that gives no detectable hydrolysis of RNA by assay on radioactive substrates.

(PDF 0-2 workdays service: $29.90)

Accession: 006173720



Related references

Preparation of immobilized monomeric actin and its use in the isolation of protease free and rnase free pancreatic dnase i. European Journal of Biochemistry 179(1): 215-220, 1989

On the preparation of protease free pancreatic dnase. Federation Proceedings 37(6): 1415, 1978

Preparation of "RNase-Free" DNase by Alkylation. Methods in Molecular Biology 2: 13-20, 1985

Small-scale preparation of RNase-free DNase. Biotechniques 6(9): 846-847, 1988

Preparation of rnase free dnase i and alpha amylase. Analytical Biochemistry 52(2): 342-348, 1973

Application of general ligand affinity chromatography for the mutual separation of dnase and rnase free of protease contamination. Analytical Biochemistry 74(1): 138-144, 1976

Preparation of immobilized monomeric actin and its use in the isolation of protease-free and ribonuclease-free pancreatic deoxyribonuclease I. European Journal of Biochemistry 179(1): 215-219, 1989

Preparation of protease-free and ribonuclease-free pancreatic deoxyribonuclease. Journal of Biological Chemistry 253(20): 7216-7219, 1978

Preparation of ribonuclease-free DNase I and -amylase. Analytical Biochemistry 52(2): 342-348, 1973

Rna synthesis in cell free extracts of barley m leaves infected with brome grass mosaic virus actinomycin d metab enz rnase enz dnase. Virology 36(2): 293-302, 1968