Merck, MABE1109, Anti-HHV-8 LNA-1 Antibody, clone LN53

General description
116/135 kDa (KSHV LNA) and 118 kDa (RFHV LNA) prediced, ~220/230 kDa doublet (KSHV) and ~240 kDa doublet (RFHV) reported (Burnside, K.L., et al. (2006). Virology. 354(1):103-115; Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155). The larger-than-predicted apparent molecular weight is most likely caused by the internal acidic repeat region sequence (Rainbow, L., et al. (1997). J. Virol. 71(8):5915-5921).
Human herpesvirus-8//Kaposi′s sarcoma-associated herpesvirus (HHV-8/KSHV) and macaque retroperitoneal fibromatosis herpesvirus (RFHV) were originally identified in in AIDS patients in association with Kaposi′s sarcoma (KS) and in retroperitoneal fibromatosis (RF) tumor lesions of macaques with simian AIDS, respectively. The most prominent protein expressed in cells latently infected with KSHV and RFHV is the orf73 gene product known as the latency-associated nuclear antigen (LANA or LNA). LANA is a nuclear protein that functions to ensure the maintenance of the viral genome by tethering the viral episomal DNA to host cell chromosomes. LANA also regulates the cellular transcription program in host cells through interaction with a number of cellular proteins, including transcriptional regulators and known tumor suppressors, p53 and pRB. In addtion, LANA directly influences the viral transcription program and helps maintain the latent state of the virus by inhibiting viral replication. KSHV LANA consists of a serine- and proline-rich N-terminal domain with a nuclear localization signal (NLS), a chromatin-binding motif (CBM), and domains responsible for interaction with the transcription regulators, mSin3 complex and Sp1. The central domain contains several highly repetitive acidic regions that vary in length and are responsible for the size variation of LANA proteins from different KSHV isolates that can range from 1003 to 1162 amino acids. The proline-rich C-terminal third domain contains another NLS and is responsible for LANA dimerization and binding to the terminal repeats (TR) of the viral genomic DNA. The C-terminal domain is responsible for interaction with tumor suppressors pRB and p53.

Immunogen
Affinity-purified GST fusions of HHV-8 LNA-1 C-terminal fragments (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Application
Anti-HHV-8 LNA-1, clone LN53, Cat. No. MABE1109, is a highly specific rat monoclonal antibody that targets human KSHV/HHV8 and simian RFHV Orf73 latency-associated nuclear antigen (LANA or LNA) and has been tested in ELISA, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Immunoprecipitati
ELISA Analysis: A representative lot detected HHV-8 LNA-1 recombinant constructs containing the repetitive region a.a. 803-929 (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Flow Cytometry Analysis: A representative lot detected HHV-8 LNA-1 immunoreactivity in BCP-1, BC-3, and HBL-6 cells, but not in Daudi or Ramos cells (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Immunocytochemistry Analysis: A representative lot detected the transiently expressed KHSV ORF73 (HHV-8 LANA or LNA-1) as well as rhesus macaque retroperitoneal fibromatosis herpesvirus ORF73 (RFHV LANA) in the nuclei of transfected COS cells by fluorescent immunocytochemistry. No reactivity was detected toward pig-tailed macaque MneRV2 or rhesus macaque RRV LANA proteins (Burnside, K.L., et al. (2006). Virology. 354(1):103-115).

Immunocytochemistry Analysis: A representative lot detected HHV-8 LNA-1-positive nuclear bodies in primary effusion lymphoma (PEL) cell line BCP-1 by fluorescent immunocytochemistry, but not in Daudi or Ramos cells (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Immunohistochemistry Analysis: A representative lot detected RFHVMn LANA immunoreactivity in the nuclei of retroperitoneal fibromatosis (RF) tumor cells by immunohistochemistry staining of formalin-fixed, paraffin-embedded RF biopsies from an SIV-infected pig-tailed macaque. No immunoreactivity was detected using normal macaque jejunum tissue sections (Burnside, K.L., et al. (2006). Virology. 354(1):103-115).

Immunohistochemistry Analysis: Representative lots immunostained cells latently infected by HHV-8 by immunohistochemistry staining of paraffin-embedded patch, plaque, and nodular Kaposi s sarcoma (KS), multicentric Castleman s disease (MCD), and primary effusion lymphoma (PEL) tissue sections (Codish, S., et al. (2000). Am. J. Hematol. 65(4):310-314; Dupin, N., et al. (2000). Blood. 95(4):1406-1412; Dupin, N., et al. (1999). Proc. Natl. Acad. Sci. U. S. A. 96(8):4546-4551).

Immunohistochemistry Analysis: A representative lot detected nuclear HHV-8 LNA-1 immunoreactivity in cells within the nodular Kaposi s sarcoma (KS) lesion, but not in the surrounding dermis by immunohistochemistry staining of paraffin-embedded classical KS tumor sections (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Immunoprecipitation Analysis: A representative lot immunoprecipitated 220/230 kDa HHV-8 LNA-1 from primary effusion lymphoma (PEL) cell line BC-3, but not from Ramos cells (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Western Blotting Analysis: A representative lot detected the transiently expressed KHSV ORF73 (HHV-8 LANA or LNA-1) as well as rhesus macaque retroperitoneal fibromatosis herpesvirus ORF73 (RFHV LANA) in lysate from transfected COS cells. No reactivity was detected toward pig-tailed macaque MneRV2 or rhesus macaque RRV LANA proteins (Burnside, K.L., et al. (2006). Virology. 354(1):103-115).

Western Blotting Analysis: A representative lot detected the 220/230 kDa HHV-8 LNA-1 doublet in lysate from primary effusion lymphoma (PEL) cell line BC-3, but not from Ramos cells (Kellam, P., et al. (1999). J. Virol. 73(6): 5149 5155).

Biochem/physiol Actions
Clone LN53 recognizes repetitive glutamic acid structural motifs EQEQE and EPEPE found in the Glu-rich repeat region of human Kaposi′s sarcoma-associated herpesvirus (KSHV or HHV-8) and simian retroperitoneal fibromatosis herpesvirus (RFHV) Orf73 latency-associated nuclear antigen (LANA or LNA), respectively.

Human KSHV (HHV8) and simian RFHV.
Physical form
Format: Purified
Purified rat monoclonal IgG2c in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide

Analysis Note
Evaluated by Immunocytochemistry in BCBL-1 cells.

Immunocytochemistry Analysis (ICC): 1 µg/mL of this antibody immunostained HHV-8 LNA-1-positive nuclear bodies in Kaposi′s sarcoma-associated herpesvirus/human herpesvirus 8- (KSHV/HHV8-) infected BCBL-1 lymphoma cells.

Other Notes
Concentration: Please refer to lot specific datasheet.
Legal Information
GenBank is a registered trademark of United States Department of Health and Human Services