Elsevier

Clinica Chimica Acta

Volume 17, Issue 2, August 1967, Pages 201-206
Clinica Chimica Acta

Glucose-6-phosphatase assay on microgram amounts of liver tissue

https://doi.org/10.1016/0009-8981(67)90119-2Get rights and content

Abstract

To make possible the assay of glucose-6-phosphatase activity on very small liver biopsy specimens a method was worked out based on the following principles: 1. Utilization of small plastic tubes, enclosed in larger glass tubes, to avoid evaporation of the small (0.02 ml) sample during several hours of incubation. 2. Stabilization of the enzyme by sucrose and EDTA. 3. Determination of liberated phosphate by a newly described, highly sensitive method, using Malachite Green.

With this method, 1.56 μg of rat liver tissue in each tube was sufficient for an assay. When more convenient, up to at least 100 μg of tissue could equally well be used.

When compared to a reference method, using 1 mg of tissue in each tube, similar values for the glucose-6-phosphatase activity were obtained. The coefficient of variation, when 20 analyses were made on the same homogenate, was 13.3%, as compared to 11.5% for the reference method.

References (5)

  • H.G. Hers et al.
  • K. Itaya et al.

    Clin. Chim. Acta

    (1966)
There are more references available in the full text version of this article.

Cited by (7)

  • Experimental evidence for the use of CCR2 antagonists in the treatment of type 2 diabetes

    2013, Metabolism: Clinical and Experimental
    Citation Excerpt :

    Hepatic glycogen content was determined with the Glycogen Assay Kit (BioVision, Mountain View, CA). Glucose-6-phosphatase (G-6-Pase) activity was determined as previously described [40] using a colorimetric assay kit (BioVision, Mountain View, CA). Hepatic G-6-Pase protein levels were analyzed by standard Western blot techniques, using a rabbit anti-G-6-Pase antibody (#sc-134714, Santa Cruz Biotechnology, Santa Cruz, CA) at a 1:300 dilution.

View all citing articles on Scopus
View full text