Immunolocalization of detergent-susceptible nucleoplasmic lamin A/C foci by a novel monoclonal antibody

Muralikrishna, Bh. ; Thanumalayan, S. ; Jagatheesan, G. ; Rangaraj, Nandini ; Karande, Anjali A. ; Parnaik, Veena K. (2004) Immunolocalization of detergent-susceptible nucleoplasmic lamin A/C foci by a novel monoclonal antibody Journal of Cellular Biochemistry, 91 (4). pp. 730-739. ISSN 0730-2312

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Official URL: http://onlinelibrary.wiley.com/doi/10.1002/jcb.107...

Related URL: http://dx.doi.org/10.1002/jcb.10765

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 C3H10T½ 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, C3H10T½ 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:Article
Source:Copyright of this article belongs to John Wiley and Sons.
Keywords:Nuclear Lamina; Nuclear Matrix; Lamin A; Nucleoplasmic Foci
ID Code:34782
Deposited On:11 Apr 2011 14:03
Last Modified:17 May 2016 17:40

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