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Title: Quantitation and characterization of rat tissue metallothioneins by gel electrophoresis

Abstract

A discontinuous gradient gel electrophoretic system was developed to quantitate and characterize metallothionein (MT) in rat tissue. Vertical slab separating gels (1.5 mm x 14 cm x 12 cm) consisted of a linear polyacrylamide gradient 7.5 to 30% T and 5% Bis. The stacking gels (3% T and 20% Bis) were photopolymerized using riboflavin as the catalyst. Liver cytosols were prepared from rats which received (i.p.) various amounts of Zn (5 mg/kg BW) or Cd (2.5 mg/kg BW). Purified MT was prepared by gel filtration and DEAE ion-exchange chromatography. Cytosols were heated (80/sup 0/C, 2 min) and centrifuged to obtain a supernatant. An appropriate amount of supernatant and various amounts of MT standard were electrophoresed (constant current, 20 mA per slab) for 9 hours. Gels were stained with Commassie Blue (R-250, 0.25%) for 12 hours and destained. Gels were scanned by densitometer and peaks heights were determined. Significantly linear standard curves (..mu..g MT vs. peak height) were established for both MTI and MTII. (Cd, Zn)-MTI migrated slower than Zn-MTI while mobilities for both (Cd, Zn)- and Zn-MTII were the same. The accumulation of MTI was consistently less than MTII in liver from both Zn- and Cd-injected rats. Their results suggestmore » that electrophoretic analysis is an excellent system not only for quantitation but also for characterization of MT in rat tissue.« less

Authors:
;
Publication Date:
Research Org.:
Cornell Univ., Ithaca, NY
OSTI Identifier:
6846599
Report Number(s):
CONF-8604222-
Journal ID: CODEN: FEPRA
Resource Type:
Conference
Journal Name:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
Additional Journal Information:
Journal Volume: 45:4; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; CADMIUM; METABOLISM; METALLOTHIONEIN; ELECTROPHORESIS; ZINC; CHEMICAL COMPOSITION; ION EXCHANGE CHROMATOGRAPHY; LIVER; RATS; ANIMALS; BODY; CHROMATOGRAPHY; DIGESTIVE SYSTEM; ELEMENTS; GLANDS; MAMMALS; METALLOPROTEINS; METALS; ORGANIC COMPOUNDS; ORGANS; PROTEINS; RODENTS; SEPARATION PROCESSES; VERTEBRATES; 560300* - Chemicals Metabolism & Toxicology

Citation Formats

Lin, L Y, and McCormick, C C. Quantitation and characterization of rat tissue metallothioneins by gel electrophoresis. United States: N. p., 1986. Web.
Lin, L Y, & McCormick, C C. Quantitation and characterization of rat tissue metallothioneins by gel electrophoresis. United States.
Lin, L Y, and McCormick, C C. Wed . "Quantitation and characterization of rat tissue metallothioneins by gel electrophoresis". United States.
@article{osti_6846599,
title = {Quantitation and characterization of rat tissue metallothioneins by gel electrophoresis},
author = {Lin, L Y and McCormick, C C},
abstractNote = {A discontinuous gradient gel electrophoretic system was developed to quantitate and characterize metallothionein (MT) in rat tissue. Vertical slab separating gels (1.5 mm x 14 cm x 12 cm) consisted of a linear polyacrylamide gradient 7.5 to 30% T and 5% Bis. The stacking gels (3% T and 20% Bis) were photopolymerized using riboflavin as the catalyst. Liver cytosols were prepared from rats which received (i.p.) various amounts of Zn (5 mg/kg BW) or Cd (2.5 mg/kg BW). Purified MT was prepared by gel filtration and DEAE ion-exchange chromatography. Cytosols were heated (80/sup 0/C, 2 min) and centrifuged to obtain a supernatant. An appropriate amount of supernatant and various amounts of MT standard were electrophoresed (constant current, 20 mA per slab) for 9 hours. Gels were stained with Commassie Blue (R-250, 0.25%) for 12 hours and destained. Gels were scanned by densitometer and peaks heights were determined. Significantly linear standard curves (..mu..g MT vs. peak height) were established for both MTI and MTII. (Cd, Zn)-MTI migrated slower than Zn-MTI while mobilities for both (Cd, Zn)- and Zn-MTII were the same. The accumulation of MTI was consistently less than MTII in liver from both Zn- and Cd-injected rats. Their results suggest that electrophoretic analysis is an excellent system not only for quantitation but also for characterization of MT in rat tissue.},
doi = {},
url = {https://www.osti.gov/biblio/6846599}, journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:4,
place = {United States},
year = {1986},
month = {3}
}

Conference:
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