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ER-ID® Red assay kit (GFP-CERTIFIED®)

Endoplasmic  Reticulum staining with minimal toxicity to cells
 
ENZ-51026-K500 1 Kit 232.00 USD
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ER-ID® Red assay kit (GFP-CERTIFIED®) contains an endoplasmic reticulum-selective dye suitable for live cell, or detergent-permeabilized aldehyde-fixed cell staining. Micromolar concentrations of ER-ID® Red 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® Red dye is in fluorescence co-localization imaging with green fluorescent protein (GFP)-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 red fluorescent dyes to green fluorescent ones and metachromatic artifacts, wherein fluorescent dyes emit both in the red and green regions of the spectrum, have led to spurious results in GFP 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®  Red dye, a new red-emitting, cell-permeable small molecule organic probe that spontaneously localizes to live or fixed endoplasmic reticula, was developed. ER-ID®  Red 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 Texas Red region of the visible light spectrum, and is highly resistant to photo-bleaching, concentration quenching and photoconversion. The ER-ID®  Red assay kit GFP-CERTIFIED®) is specifically designed for use with GFP-expressing cell lines, as well as cells expressing blue, cyan or yellow fluorescent proteins (BFPs, CFPs, YFPs). 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 fluorescein, or coumarin. A nuclear counterstain is provided to highlight this organelle as well.
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Figure 1: Live HeLa cells stained with ER-ID® Red dye (A), Hoechst dye (B) and resulting composite image (C).
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Product Specification

Applications:Fluorescence microscopy
 
Application Notes:Enzo Life Sciences’ ER-ID® Red assay kit (GFP-CERTIFIED®) contains a novel endoplasmic reticulum-selective dye suitable for live cell, or detergent-permeabilized aldehyde-fixed cell staining.
 
Quality Control:A sample from each lot of ER-ID® Red assay kit is used to stain HeLa cells, using the procedures described in the user manual. The selectivity of the ER-ID® Red dye is evident.
 
Quantity:500 assays
 
Use/Stability:With proper storage, the kit components are stable up to the date noted on the product label. Store kit at -20°C in a non-frost free freezer, or -80°C for longer term storage.
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Kit/Set Contains:ER-ID® Red detection reagent: 50μl
Hoechst 33342 nuclear stain: 50μl
10X assay buffer: 15ml
 
Technical Info/Product Notes:The ER-ID® Red assay kit (GFP-CERTIFIED®) is a member of the CELLESTIAL® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLESTIAL® reagents and kits are optimal for use in demanding cell analysis applications involving confocal microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.
 

Product Literature References

AT-101 simultaneously triggers apoptosis and a cytoprotective type of autophagy irrespective of expression levels and the subcellular localisation of Bcl-xL and Bcl-2 in MCF7 cells : P. Antonietti, et al. ; Biochim. Biophys. Acta 1863, 499 (2016), Application(s): Fluorescence microscopy, Abstract;
Domain architecture of vasohibins required for their chaperone-dependent unconventional extracellular release: T. Kadonosono, et al.; Protein Sci. (2016), Abstract;
Biogenesis of the crystalloid organelle in Plasmodium involves microtubule-dependent vesicle transport and assembly: S. Saeed, et al.; Int. J. Parasitol. 45, 537 (2015), Application(s): Microscopy, Abstract;
Synthesis of a peptide that can translocate to the endoplasmic reticulum: K. Nakatsu, et al.; Biochem. Biophys. Res. Commun. 460, 328 (2015), Application(s): Fluorescent Assay, Abstract;
Target identification of grape seed extract in colorectal cancer using drug affinity responsive target stability (DARTS) technique: role of endoplasmic reticulum stress response proteins: M.M. Derry, et al.; Curr. Cancer Drug Targets 14, 323 (2014), Abstract;
Targeted protein destabilization reveals an estrogen-mediated ER stress response: K. Raina, et al.; Nat. Chem. Biol. 10, 957 (2014), Application(s): Confocal microscopy, Abstract; Full Text
Enhanced optineurin E50K-TBK1 interaction evokes protein insolubility and initiates familial primary open-angle glaucoma: Y. Minegishi, et al.; Hum. Mol. Genet. 22, 3559 (2013), Abstract;
Knockdown of RyR3 enhances adiponectin expression through an atf3-dependent pathway: S.H. Tsai, et al.; Endocrinology 154, 1117 (2013), Abstract;
Molecular cloning and tumour suppressor function analysis of canine REIC/Dkk-3 in mammary gland tumors: K. Ochiai, et al.; Vet. J. 197, 769 (2013), Application(s): ER staining in dog mammary carcinoma cells, Abstract;
Snowflake vitreoretinal degeneration (SVD) mutation R162W provides new insights into Kir7.1 ion channel structure and function: B.R. Pattnaik, et al.; PLoS One 8, e71744 (2013), Application(s): ER staining in CHO-K1 cells, Abstract; Full Text

General Literature References

Photoconversion of Lysotracker Red to a green fluorescent molecule: E.C. Freundt, et al.; Cell Res. 17, 956 (2007), Abstract;
Systematic colocalization errors between acridine orange and EGFP in astrocyte vesicular organelles: F. Nadrigny, et al.; Biophys. J. 93, 969 (2007), Abstract;
Chloromethyl-X-rosamine (MitoTracker Red) photosensitises mitochondria and induces apoptosis in intact human cells: T. Minamikawa, et al.; J. Cell Sci. 112, 2419 (1999), Abstract;
Chloromethyltetramethylrosamine (Mitotracker Orange) induces the mitochondrial permeability transition and inhibits respiratory complex I. Implications for the mechanism of cytochrome c release: L. Scorrano, et al.; J. Biol. Chem. 274, 24657 (1999), Abstract;

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