Preparation and characterization of monoclonal antibody against protein tyrosine phosphatase PRL-3.
PRL-3 is a newly identified nonclassical protein tyrosine phosphatase associated with colorectal cancer metastasis. To prepare specific monoclonal antibody (MAb) against PRL-3, cDNAs of PRL-1, -2, and -3 were cloned by RT-PCR and inserted into prokaryotic expression vector pGEX-4T1, respectively. The fusion proteins were expressed in Escherichia coli. Monoclonal antibody against PRL-3 was obtained with hybridoma technique and specific ELISA screening. Western blotting and immunohistochemistry assays showed that MAb 3B6 had specific binding ability with PRL-3 protein in eukaryotic cells. This MAb will be a helpful tool for the detection of PRL-3 protein in the tissues and serum of colorectal cancer and other cancer patients.
Department of Biochemistry and Molecular Biology, Beijing Institute for Cancer Research, Peking University School of Oncology, Beijing, People's Republic of China.
Lirong
L
Peng
Yan
Y
Li
Ling
L
Meng
Chengchao
C
Shou
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay; D019073:HT29 Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology*; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000276:immunology*; D011995:Recombination, Genetic
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D018076:DNA, Complementary; D017874:Immediate-Early Proteins; D009363:Neoplasm Proteins; C484730:PTP4A3 protein, human; D017027:Protein Tyrosine Phosphatases