|
|
|
|
 |
|||
|
|||
Volume 52 (2): 253-261, 2004
Copyright ©The Histochemical Society, Inc.
Generation and Characterization of Anti-MUC4 Monoclonal Antibodies Reactive with Normal and Cancer Cells in Humans
Department of Biochemistry and Molecular Biology and Eppley Institute for Research in Cancer and Allied Disease, University of Nebraska Medical Center, Omaha, Nebraska (NM,GCV,SCC,MJ,UAW,MA,SKB); Institute of Microbial Technology, Chandigarh, India (GCV); and Unité INSERM 560, Lille, France (MCC,JPA)
Correspondence to: Surinder K. Batra, PhD, Dept. of Biochemistry and Molecular Biology, Eppley Inst. for Research in Cancer and Allied Diseases, U. of Nebraska Medical Center, 984525 Nebraska Medical Center, Omaha, NE 68198–4525. E-mail: sbatra{at}unmc.edu
We have previously cloned the full-length cDNA (approximately 28 Kb) and established the complete genomic organization (25 exons/introns over 100 kb) of the human MUC4 mucin. This large molecule is predicted to protrude over 2 µm above the cell surface, in which MUC4
is an extracellular mucin-type glycoprotein subunit and MUC4ß is the transmembrane subunit. Over two thirds of the encoded protein sequence consists of 16-amino-acid tandem repeats (TR), which are flanked by unique sequences. In this study we generated and characterized monoclonal antibodies (MAbs) directed against the TR region of MUC4. Mice were immunized with a KLH-conjugated MUC4 TR peptide, STGDTTPLPVTDTSSV. Several clones were purified by three rounds of limited dilutions and stable clones presenting a sustained antibody production were selected for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the MUC4 peptide and further screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses. One of the MAbs (8G7) was strongly reactive against the MUC4 peptide and with native MUC4 from human tissues or pancreatic cancer cells in Western blotting, immunohistochemistry, and confocal analysis. Anti-MUC4 MAb may represent a powerful tool for the study of MUC4 function under normal and pathological conditions and for diagnosis of solid tumors including those in the breast, pancreas, lungs, and ovaries.
(J Histochem Cytochem 52:253–261, 2004)
Key Words: mucin • MUC4 • antibody • pancreatic adenocarcinoma
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Vincent, M.-P. Ducourouble, and I. Van Seuningen Epigenetic regulation of the human mucin gene MUC4 in epithelial cancer cell lines involves both DNA methylation and histone modifications mediated by DNA methyltransferases and histone deacetylases FASEB J, August 1, 2008; 22(8): 3035 - 3045. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. D. Blalock, S. J. Spurr-Michaud, A. S. Tisdale, and I. K. Gipson Release of Membrane-Associated Mucins from Ocular Surface Epithelia Invest. Ophthalmol. Vis. Sci., May 1, 2008; 49(5): 1864 - 1871. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Chaturvedi, A. P. Singh, S. Chakraborty, S. C. Chauhan, S. Bafna, J. L. Meza, P. K. Singh, M. A. Hollingsworth, P. P. Mehta, and S. K. Batra MUC4 Mucin Interacts with and Stabilizes the HER2 Oncoprotein in Human Pancreatic Cancer Cells Cancer Res., April 1, 2008; 68(7): 2065 - 2070. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Chaturvedi, A. P. Singh, N. Moniaux, S. Senapati, S. Chakraborty, J. L. Meza, and S. K. Batra MUC4 Mucin Potentiates Pancreatic Tumor Cell Proliferation, Survival, and Invasive Properties and Interferes with Its Interaction to Extracellular Matrix Proteins Mol. Cancer Res., April 1, 2007; 5(4): 309 - 320. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. L. Russo, S. Spurr-Michaud, A. Tisdale, J. Pudney, D. Anderson, and I. K. Gipson Mucin gene expression in human male urogenital tract epithelia Hum. Reprod., November 1, 2006; 21(11): 2783 - 2793. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Davidson, Z. Zhang, L. Kleinberg, M. Li, V. A. Florenes, T.-L. Wang, and I.-M. Shih Gene expression signatures differentiate ovarian/peritoneal serous carcinoma from diffuse malignant peritoneal mesothelioma. Clin. Cancer Res., October 15, 2006; 12(20): 5944 - 5950. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Koscinski, S. Viville, N. Porchet, A. Bernigaud, F. Escande, A. Defossez, and M.-P. Buisine MUC4 gene polymorphism and expression in women with implantation failure Hum. Reprod., September 1, 2006; 21(9): 2238 - 2245. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Moniaux, W. M. Junker, A. P. Singh, A. M. Jones, and S. K. Batra Characterization of Human Mucin MUC17: COMPLETE CODING SEQUENCE AND ORGANIZATION J. Biol. Chem., August 18, 2006; 281(33): 23676 - 23685. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |