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http://krishi.icar.gov.in/jspui/handle/123456789/4881
Title: | Monoclonal antibody to serum immunoglobulins of Clarias batrachus and its application in immunoassays |
Other Titles: | Not Available |
Authors: | N. Sood D.K. Chaudhary A. Singh G. Rathore |
Published/ Complete Date: | 2012-09-19 |
Project Code: | Not Available |
Keywords: | Clarias batrachus Flow cytometry ELISA Immunoglobulin Immunoperoxidase test Monoclonal antibody |
Publisher: | ELSEVIER |
Citation: | Sood N, Chaudhary DK, Singh A, Rathore G. Monoclonal antibody to serum immunoglobulins of Clarias batrachus and its application in immunoassays. Gene. 2012 Dec 15;511(2):411-9. doi: 10.1016/j.gene.2012.09.044. Epub 2012 Sep 19. PubMed PMID: 23000018. |
Series/Report no.: | Not Available; |
Abstract/Description: | Serum immunoglobulins of Clarias batrachus (Cb-Ig) were purified by affinity chromatography using bovine serum albumin as capture ligand. Under reducing conditions in SDS-PAGE, Cb-Ig was composed of a heavy (H) chain (68.7 kDa) and two light (L) chains (27.4 and 26.3 kDa). Purified Cb-Ig was used to produce a monoclonal antibody (MAb) designated E4 MAb that belonged to IgG1 subclass. In Western blotting, this MAb showed binding to H chain of purified Cb-Ig and putative H chains in reduced sera of C. batrachus, Clarias gariepinus and Heteropneustes fossilis. However, no binding was observed with serum protein of Labeo rohita and Channa striata. Cross-reactivity of anti-Cb-Ig MAb was observed with serum of C. batrachus, C. gariepinus and H. fossilis in competitive ELISA. In immunoblotting of non-reduced Cb-Ig with E4 MAb, four bands assumed to be tetrameric, trimeric, dimeric and monomeric form were observed. In flow cytometric analysis of the gated lymphocytes, the number of surface Ig-positive (Ig +) cells in blood, spleen, kidney and thymus of C. batrachus was determined to be 50.1 ± 3.1, 55.1 ± 3.36, 42.4 ± 4.81 and 5.1 ± 0.89%, respectively, using E4 MAb. Ig + cells were also demonstrated in formalin-fixed paraffin embedded tissue sections of spleen, kidney, thymus and smears of blood mononuclear cells in indirect immunoperoxidase test. The developed MAb was employed to detect pathogen-specific immunoglobulins in the sera of C. batrachus immunized with killed Edwardsiella tarda, by an indirect ELISA. This monoclonal antibody can be useful tool in immunological research and assays. |
Description: | Not Available |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | Gene |
NAAS Rating: | 8.98 |
Volume No.: | 511 |
Page Number: | 411-419 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | http://dx.doi.org/10.1016/j.gene.2012.09.044 |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/4881 |
Appears in Collections: | FS-NBFGR-Publication |
Files in This Item:
File | Description | Size | Format | |
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1-s2.0-S0378111912011250-main.pdf | 1.18 MB | Adobe PDF | View/Open |
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