| MaxLight(TM) 650 is a new Far-IR stable dye conjugate comparable to Alexa Fluor(TM)647, DyLight(TM)649, Cy5(TM) and offers better labeling efficiency, brighter imaging and increased immunodetection. Absorbance (655nm), Emission (676nm), Extinction Coefficient 250,000. ZO-3, a ~130kD protein, contains three PSD-95/SAP90, discs-large, ZO-1 (PDZ) domains, a SH3 domain, and a guanylate kinase-liked domain. Like ZO-1 and ZO-2, ZO-3 contains a C-terminal acidic domain and a basic region (between the first and second PDZ domains), making it belongs to the MAGUK protein family. By Western Blot analysis, ZO-3 is identified as a ~130kD protein that directly interacts with tight junction proteins ZO-1 and occuldin, but not with ZO-2. However, in one study, ZO-2, ZO-3 and Occludin colocalized with actin aggregates at cell border in MDCK cells. ZO-3 is localized to sites of cell-cell interaction, identical to that of ZO-1 in tight junctions. By Immunoprecipitation analysis, ZO-3 co-immunoprecipitates with ZO-1. Using MDCK cells, ZO-3 protein colocalized with ZO-1 by Immunofluorescence. In vitro binding assays, the N-terminal of ZO-3 showed preferably binding of ZO-1 and actin filaments. Moreover, the binding of ZO-1/ZO-2/ZO-3 was also found to interact with certain Claudin proteins. Applications: Suitable for use in Immunofluorescence, FLISA, Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilutions: Immunofluorescence: 0.5-2ug/ml FLISA: 0.1-1ug/ml Western Blot: 1-3ug/ml Immunoprecipitation: 2-3ug Optimal dilutions to be determined by the researcher. Storage and Stability: Store product at 4C in the dark. DO NOT FREEZE! Stable at 4C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight(TM)650 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Note: Applications are based on unconjugated antibody. |
