Paraffin Sections (P), Enzyme Immunoassay (E), Immunocytochemistry/Immunofluorescence (ICC/IF), Western blot / Immunoblot (WB), Frozen Sections (C), Immunoprecipitation (IP)
Background of Herpes Virus Type 8 / HHV8 antibody
ORF57 (also known as MTA), one of the earliest Kaposi's sarcoma-associated herpesvirus (KSHV) regulatory proteins to be expressed, is essential for virus lytic replication. A counterpart is present in every herpesvirus sequenced, indicating the importance of this signature viral protein, and those examined act post-transcriptionally, affecting RNA splicing and transport. KSHV ORF57 is capable of establishing both lytic and latent replication cycles. In KS, the virus localizes to tumor progenitor endothelial cells, most of which are latently infected. In cell culture, KSHV replication is generally studied using B-cell lines, such as BCBL-1, generated from primary effusion lymphoma material. Most BCBL-1 cells are latently infected, although there is some spontaneous virus reactivation. Addition of chemical inducers such as sodium n-butyrate, 12-O-tetradecanoylphorbol-13-acetate (TPA), and valproic acid (VA) to these cells efficiently induces the lytic cycle and produces virions. KSHV ORF57 protein predominantly localizes to the nucleus and can shuttle between the nucleus and cytoplasm. Most HSV-1 genes are unspliced; by contrast, ORF57 is spliced gene; the protein is 455 amino acids in length and 50kDa in size.
Cultured cells infected with HHV8 stained with HHV8 antibody DM395 /DM395-05 (Clone LN53).
Immunohistochemistry-Paraffin: KSHV K8 Antibody (8C12G10G1) [NB100-2189] - Immunohistochemical analysis of paraffin-embedded lung cancer tumor (left), mammary cancer tissues (right) using c-Jun Rabbit pAb with DAB staining.
Western Blot: KSHV ORF8 Antibody [NB100-2197] - Western blot analysis using anti-KSHV ORF8 polyclonal antiobdy against uninduced BCBL1 cell lysate (1), TPA induced BCBL1 cell lysate(2) and purified virion (3).
Western blot using HHV8gp59 polyclonal antibody ( Cat # PAB10254 ) to detect HHV8gp59 in HEK293 cells transfected with HHV8gp59 expression vector and HHV8gp59 truncations, or in HHV8gp59 infected B-cell line ( BCBL-1 ) treated with or without valproic acid to induce viral replication ( arrow ).The membrane was probed with the primary antibody diluted 1 : 7,500 ( left ) and 1:1,000 ( right ).Personal Communication, V. Majerciak, M.Zheng, CCR-NC , Bethesda, MD.
Transfected and non-transfected 293T cell lysates were resolved by SDS-PAGE, transferred to NC membrane and probed with ORF73/HHV8 monoclonal antibody, clone AT4C11 ( Cat # MAB4201 ; 1 : 1,000 ). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.
Immunocytochemical staining of TPA-induced BCBL-1 cells and uninduced BCBL-1 cells, using KSHV K8a monoclonal antibody, clone 8C12G10G1 ( Cat # MAB5592 ).HRP-anti-mouse was used as the secondary antibody before color development with AEC.
Western blot using affinity purified anti-KSHV FORF57 to detect KSHV ORF57 in HEK293 cells transfected with ORF57 expression vector and ORF57 truncations, or in KSHV infected B-cell line (BCBL-1) treated with or without valproic acid to induce viral replication (arrow). The membrane was probed with the primary antibody diluted 1:7,500 (left) and 1:1,000 (right).