Enzo Life Sciences’ ER-ID™ Green assay kit contains a novel endoplasmic reticulum-selective dye suitable for live cell, or detergent-permeabilized aldehyde-fixed cell staining. Micromolar concentrations of ER-ID™ Green dye are sufficient for staining mammalian cells, as validated with human cervical carcinoma cell line, human T-lymphocyte cell line, Jurkat, HeLa and human bone osteosarcoma epithelial cell line, U2OS.One important application of ER-ID™ Green dye is in fluorescence co-localization imaging with red fluorescent protein (RFP)- or orange fluorescent protein (OFP)-tagged proteins, a powerful approach for determining the targeting of molecules to intracellular compartments and for screening of their associations and interactions. However, to date, photoconversion of fluorescent dyes to other colors and metachromatic artifacts, wherein fluorescent dyes emit both in the red (or orange) and green regions of the spectrum, have led to spurious results in co-localization experiments. Additionally, many organelle-targeting probes photobleach rapidly, are subject to quenching upon concentration in organelles, are highly toxic, or only transiently associate with the target organelle, requiring imaging within a minute or two of dye addition. Consequently, ER-ID™ Green dye, a new green-emitting, cell-permeable small molecule organic probe that spontaneously localizes to live or fixed endoplasmic reticula, was developed. ER-ID™ Green dye can be readily used in combination with other common UV and visible light excitable organic fluorescent dyes and various fluorescent proteins in multi-color imaging and detection applications. The dye emits in the FITC region of the visible light spectrum, and is resistant to photo-bleaching, concentration quenching and photoconversion.The ER-ID™ Green assay kit is specifically designed for use with RFP- or OFP-expressing cell lines, as well as cells expressing blue or cyan fluorescent proteins (BFPs, CFPs). Additionally, the kit is suitable for use with live or post-fixed cells in conjunction with probes, such as labeled antibodies, or other fluorescent conjugates displaying similar spectral properties as Texas Red, or coumarin. A nuclear counterstain is provided to highlight this organelle as well.

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