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.
Peng
Lirong
L
Department of Biochemistry and Molecular Biology, Beijing Institute for Cancer Research, Peking University School of Oncology, Beijing, People's Republic of China.
Li
Yan
Y
Meng
Ling
L
Shou
Chengchao
C
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