Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting
Background: Kinetochores are mitotic structures that form on centromeres and attach to mitotic spindle microtubules. Kinetochore attachment to microtubules regulates chromosome segregation and progression through mitosis. Unattached kinetochores signal to the spindle assembly checkpoint (SAC) machinery, arresting cells in mitosis (1). CASC5, also known as Knl1 or Blinkin, is the largest subunit of the Knl1–Mis12–Ndc80 complex (KMN) network, a structural component of kinetochores required for microtubule binding. CASC5 functions in the formation of kinetochore–microtubule attachments, chromosome segregation, and in activating the SAC. CASC5 has been implicated in human diseases, including leukemia and microcephaly (2). Activation of the SAC is regulated in part by mitotic phosphorylation of CASC5 at several sites, including Ser24, Ser60, Thr943, and Thr1155 (3, 4). The sequences surrounding Thr943 and Thr1155 are identical.
Application Methods: Immunohistochemistry (Paraffin), Western Blotting
Background: Nectin-2, also known as CD112 and poliovirus receptor-related 2 (PVRL2), is a single-pass type I membrane glycoprotein ubiquitously expressed on various human tissues (1). It is a calcium independent cell adhesion molecule known to interact with several cell surface receptors, including DNAM-1 (CD226), LFA-1 (CD11a), Nectin-3 (CD113), TIGIT (VSTM3), and PVRIG (CD112R) (2-7). It is one of the major constituents of adherins junctions, and therefore plays a central role in a number of cellular processes, including adhesion, migration, and proliferation (2-8). Within the immune system, Nectin-2 modulates immune cell signaling. Upon interaction with DNAM-1 expressed on T and NK cells, Nectin-2 stimulates proliferation and cytokine production (4). Upon interaction with PVRIG, Nectin-2 inhibits proliferation (7). Thus, Nectin-2 can be either a co-stimulator or a co-inhibitor of immune cell function depending on competitive receptor interactions. Nectin-2 also serves as an entry for certain mutant strains of herpes simplex virus and pseudorabies virus, and it is involved in cell to cell spreading of these viruses (8,9). Alternate transcriptional splice variants, encoding different isoforms, have been characterized (10,11).