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Immunolocalization of Detergent-Susceptible Nucleoplasmic Lamin A/C Foci by a Novel Monoclonal Antibody

Muralikrishna, BH and Thanumalayan, S and Jagatheesan, G and Rangaraj, Nandini and Karande, Anjali A and Parnaik, Veena K (2004) Immunolocalization of Detergent-Susceptible Nucleoplasmic Lamin A/C Foci by a Novel Monoclonal Antibody. In: Journal of Cellular Biochemistry, 91 (4). pp. 730-739.

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Abstract

The A-type lamins are localized in the interior of the nucleus as well as on the nuclear periphery. In this study, we have characterized a monoclonal antibody LA-2F9 produced against recombinant rat lamin A which stains a subpopulation of various cell types in a pattern of small nucleoplasmic foci that are unusually susceptible to mild detergent/salt extraction. The specific reactivity of mAb LA-2F9 towards lamins was confirmed by immunoblotting of HeLa and C3H10T1/2 whole cell lysates and nuclear lysates. The epitope for LA-2F9 was narrowed down to amino acid residues 268-278 (SAKLDNARQSA). To check whether the appearance of lamin foci was cell-cycle-dependent, C3H10T1/2 cells were serum-starved and then refed to trigger cells to enter the G1 phase of the cell-cycle. The intensity of staining increased 3.5-fold within 6 h of refeeding, when the maximum number of cells were labeled with LA-2F9. We also checked whether the LA-2F9 foci colocalized with nuclear proteins known to be distributed in small foci such as hnRNPs, snRNPs, SC-35, and p80 coilin, but did not find evidence of colocalization. Our studies suggest that LA-2F9 has a novel and specific reactivity towards detergent-susceptible lower order lamin structures that are likely to be assembly intermediates.

Item Type: Journal Article
Additional Information: The copyright belongs to Wiley-Liss, Inc.
Keywords: nuclear lamina;nuclear matrix;lamin a;nucleoplasmic foci
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 13 Dec 2005
Last Modified: 19 Sep 2010 04:21
URI: http://eprints.iisc.ernet.in/id/eprint/4295

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