CloneID: 37-5
Antigen Long Description: The original antibody was generated by immunizing Wistar rats with a mammalian expression vector encoding hOCLN.
Buffer Composition: PBS with 0.02% Proclin 300.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: Q16625
Specificity Statement: This antibody is specific for the first extracellular loop (EL1) of OCLN.
Application Notes (Clone): The cell binding capability of the original format of this antibody (rat IgG2b) was assessed using FC, and its binding to HT1080/hOCLN cells was assessed using a cell-based ELISA. This antibody was found to have no negative effect on tight junction (TJ) barrier function, as determined by a TJ barrier function assay (TER). Furthermore, this antibody was found to have no negative effect on cell viability, as determined by a colorimetric XTT assay. This antibody was found to strongly inhibit (in vitro) and fully block (in vivo) HCV infections, as measured by ICC (Shimizu et al., 2018; PMID: 29437969). This antibody's Kd for hOCLN is 0.720 ± 0.084 nM, as measured in a cell-based competition ELISA. This antibody successfully precipitated FLAG-hOCLN proteins found in HT1080 cell lysates and endogenously expressed cellular OCLN found in Caco-2, HepG2, and MCF-7 cell lysates. Furthermore, other TJ proteins expressed in T84 cells — TRIC and ZO-2 — were found to be co-immunoprecipitated with OCLN by this antibody. This antibody can be used for immunoblot analysis of OCLN (only at high concentrations due to a weak response). Immunofluorescence (IF) staining of OCLN using clone 1-3 with a TJ marker protein ZO-1 in Caco-2 cells was co-localized with ZO-1 at cell-cell contact sites. This antibody cross-reacts with crab-eating monkey OCLN (moOCLN), as determined by FC (Shimizu et al., 2019; PMID: 31077306).
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