TY - JOUR
T1 - Antiserum specific for the intact isoform-3 of metallothionein
AU - Tokheim, Abigail M.
AU - Armitage, Ian M
AU - Martin, Bruce L.
PY - 2005/4/29
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
UR - http://www.scopus.com/inward/record.url?scp=17744372812&partnerID=8YFLogxK
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U2 - 10.1016/j.jbbm.2005.01.003
DO - 10.1016/j.jbbm.2005.01.003
M3 - Article
C2 - 15892977
AN - SCOPUS:17744372812
SN - 0165-022X
VL - 63
SP - 43
EP - 52
JO - Journal of Biochemical and Biophysical Methods
JF - Journal of Biochemical and Biophysical Methods
IS - 1
ER -