Antiserum specific for the intact isoform-3 of metallothionein

Abigail M. Tokheim; Ian M Armitage; Bruce L. Martin

doi:10.1016/j.jbbm.2005.01.003

Antiserum specific for the intact isoform-3 of metallothionein

Abigail M. Tokheim, Ian M Armitage, Bruce L. Martin

Chemical and Structural Biology (TMED)

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

The recombinant form of isoform-3 of mouse brain metallothionein (MT3) was used as an antigen to immunize rabbits and raise MT3-selective antiserum. The antiserum was essentially specific for MT3 with 100-fold greater sensitivity for MT3 compared to MT1 or MT2. Immunonblot analysis of whole mouse brain homogenates showed that MT3 was present only in the fraction retained by a 30,000-Da cut-off filter. The antiserum was used to immunoprecipitate MT3 from mouse brain extracts of Swiss Webster mice and provided evidence that MT3 was a member of a macromolecular complex of greater than 30,000 Da mass in brain. An ELISA was developed using purified, recombinant mouse brain Cd₇-MT3 as the antigen and used to quantify MT3 in mouse brain extracts. The concentration of MT3 was found to be 3.0 ± 0.8 μg/ml or approximately 3.5 μg/g mouse brain (wet weight).

Original language	English (US)
Pages (from-to)	43-52
Number of pages	10
Journal	Journal of Biochemical and Biophysical Methods
Volume	63
Issue number	1
DOIs	https://doi.org/10.1016/j.jbbm.2005.01.003
State	Published - Apr 29 2005

Keywords

Metallothionein-3, antiserum
Metallothionein-3, neuronal isoform

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10.1016/j.jbbm.2005.01.003

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title = "Antiserum specific for the intact isoform-3 of metallothionein",

abstract = "The recombinant form of isoform-3 of mouse brain metallothionein (MT3) was used as an antigen to immunize rabbits and raise MT3-selective antiserum. The antiserum was essentially specific for MT3 with 100-fold greater sensitivity for MT3 compared to MT1 or MT2. Immunonblot analysis of whole mouse brain homogenates showed that MT3 was present only in the fraction retained by a 30,000-Da cut-off filter. The antiserum was used to immunoprecipitate MT3 from mouse brain extracts of Swiss Webster mice and provided evidence that MT3 was a member of a macromolecular complex of greater than 30,000 Da mass in brain. An ELISA was developed using purified, recombinant mouse brain Cd7-MT3 as the antigen and used to quantify MT3 in mouse brain extracts. The concentration of MT3 was found to be 3.0 ± 0.8 μg/ml or approximately 3.5 μg/g mouse brain (wet weight).",

keywords = "Metallothionein-3, antiserum, Metallothionein-3, neuronal isoform",

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AU - Tokheim, Abigail M.

AU - Armitage, Ian M

AU - Martin, Bruce L.

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Y1 - 2005/4/29

N2 - The recombinant form of isoform-3 of mouse brain metallothionein (MT3) was used as an antigen to immunize rabbits and raise MT3-selective antiserum. The antiserum was essentially specific for MT3 with 100-fold greater sensitivity for MT3 compared to MT1 or MT2. Immunonblot analysis of whole mouse brain homogenates showed that MT3 was present only in the fraction retained by a 30,000-Da cut-off filter. The antiserum was used to immunoprecipitate MT3 from mouse brain extracts of Swiss Webster mice and provided evidence that MT3 was a member of a macromolecular complex of greater than 30,000 Da mass in brain. An ELISA was developed using purified, recombinant mouse brain Cd7-MT3 as the antigen and used to quantify MT3 in mouse brain extracts. The concentration of MT3 was found to be 3.0 ± 0.8 μg/ml or approximately 3.5 μg/g mouse brain (wet weight).

AB - The recombinant form of isoform-3 of mouse brain metallothionein (MT3) was used as an antigen to immunize rabbits and raise MT3-selective antiserum. The antiserum was essentially specific for MT3 with 100-fold greater sensitivity for MT3 compared to MT1 or MT2. Immunonblot analysis of whole mouse brain homogenates showed that MT3 was present only in the fraction retained by a 30,000-Da cut-off filter. The antiserum was used to immunoprecipitate MT3 from mouse brain extracts of Swiss Webster mice and provided evidence that MT3 was a member of a macromolecular complex of greater than 30,000 Da mass in brain. An ELISA was developed using purified, recombinant mouse brain Cd7-MT3 as the antigen and used to quantify MT3 in mouse brain extracts. The concentration of MT3 was found to be 3.0 ± 0.8 μg/ml or approximately 3.5 μg/g mouse brain (wet weight).

KW - Metallothionein-3, antiserum

KW - Metallothionein-3, neuronal isoform

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JO - Journal of Biochemical and Biophysical Methods

JF - Journal of Biochemical and Biophysical Methods

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ER -

Antiserum specific for the intact isoform-3 of metallothionein

Abstract

Keywords

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