Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1252933 | Chemical Research in Chinese Universities | 2008 | 5 Pages |
Abstract
This study is focused on the expression of an SH2 domain-truncated form of protein tyrosine phosphatase SHP-1 (designated ÎSHP-1) and the preparation of its polyclonal antibodies. A cDNA fragment encoding ÎSHP-1 was amplified by PCR and then cloned into the pT7 expression vector. The recombinant pT7-ÎSHP-1 plasmid was used to transform Rosetta(DE3) E. coli cells. ÎSHP-1 was distributed in the exclusion body of E. coli cell extracts and was purified through a two-column chromatographic procedure. The purified enzyme exhibited an expected molecular weight on SDS-gels and HPLC gel filtration columns. It possesses robust tyrosine phosphatase activity and shows typical enzymatic characteristics of classic tyrosine phosphatases. To generate polyclonal anti-ÎSHP-1 antibodies, purified recombinant ÎSHP-1 was used to immunize a rabbit. The resultant anti-serum was subjected to purification on ÎSHP-1 antigen affinity chromatography. The purified polyclonal antibody displayed a high sensitivity and specificity toward ÎSHP-1. This study thus provides the essential materials for further investigating the biological function and pathological implication of SHP-1 and screening the inhibitors and activators of the enzyme for therapeutic drug development.
Related Topics
Physical Sciences and Engineering
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Chemistry (General)
Authors
Wan-nan LI, Yan ZHUANG, He LI, Ying SUN, Yao FU, Xiao-xia WU, Zhi-zhuang ZHAO, Xue-qi FU,