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GOLGI ID® Green assay kit

Dependably  localizes to Golgi apparatus with minimal staining of ER
 
ENZ-51028-K100 1 Kit 231.00 USD
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  • Novel Golgi apparatus-selective dye stains live or aldehyde-fixed cells
  • Fluorescence emission is easily multiplexed with common fluorescent dyes and fluorescent proteins
  • Highly resistant to photo-bleaching, concentration quenching and photo-conversion
  • Stringently manufactured to control and eliminate non-specific assay artifacts
The GOLGI ID® Green assay kit contains a Golgi apparatus-selective dye suitable for live cell, or aldehyde-fixed cell staining. Compared with other commercially available dyes for labeling Golgi bodies, GOLGI ID® Green dye is more faithfully localized to the target organelle, with minimal staining of the endoplasmic reticulum. Micromolar concentrations of GOLGI ID® Green dye are sufficient for staining mammalian cells, as validated with human cervical carcinoma cell line HeLa, human T-lymphocyte cell line, Jurkat, canine kidney cell line MDCK, and human bone osteosarcoma epithelial cell line U2OS. One important application of GOLGI ID® Green dye is in fluorescence co-localization imaging with red fluorescent protein (RFP)-tagged proteins. This is a powerful approach for determining the targeting of molecules to intracellular compartments and for screening of their associations and interactions. Many organelle-targeting probes photo-bleach 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, GOLGI ID® Green dye, a new green-emitting, cell-permeable small molecule organic probe that spontaneously localizes to live or fixed Golgi apparatus, was developed. GOLGI 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 multicolor 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 photo-conversion. The kit should also be suitable for identifying Golgi body perturbing agents and thus can be a useful tool for examining retrograde flow mechanisms in cellular secretory pathways. The GOLGI ID® Green assay kit has been specifically designed for use with RFP-expressing cell lines as well as cells expressing blue, cyan or orange fluorescent proteins (BFPs, CFPs, OFPs). Additionally, the kit is suitable for use with live or fixed cells in conjunction with probes, such as labeled antibodies, or other fluorescent conjugates displaying similar spectral properties as Texas Red, or coumarin. A blue nuclear counterstain, Hoechst 33342, is provided to highlight this organelle as well.
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Figure 1: GOLGI-ID® Green dye co-localizes with N-acetylgalactosaminyltransferase-2, fused to red fluorescent protein (RFP). (A) GOLGI-ID® Green dye, (B) HeLa cells expressing RFP-N-acetylgalactosaminyltransferase-2, (C) Hoechst 33342 staining, (D) composite image.
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Product Specification

Applications:Fluorescence microscopy
 
Application Notes:The GOLGI ID® Green assay kit is specifically designed for use in the fluorescence co-localization imaging of red fluorescent protein (RFP)-expressing cell lines as well as cells expressing blue, cyan or orange fluorescent proteins (BFPs, CFPs, OFPs), a powerful approach for determining the targeting of molecules to intracellular compartments and for screening of their associations and interactions. The kit contains a Golgi apparatus-selective dye suitable for live cell, or aldehyde-fixed cell staining.
 
Quality Control:A sample from each lot of GOLGI ID® Green assay kit is used to stain HeLa cells, using the procedures described in the user manual. The selectivities of the GOLGI ID® Green dye and the Hoechst 33342 dye are evident.
 
Quantity:100 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:GOLGI ID® Green detection reagent: 50nmoles
Hoechst 33342 nuclear stain: 50μl
10X assay buffer 1: 15ml
50X assay buffer 2: 1.2ml
 
Technical Info/Product Notes:The GOLGI ID® Green assay kit 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

Identification of a novel glycan processing enzyme with exo-acting β-allosidase activity in the Golgi apparatus using a new platform for the synthesis of fluorescent substrates: W. Hakamata, et al.; Bioorg. Med. Chem. 23, 73 (2015), Abstract;
Localization-dependent cell-killing effects of protoporphyrin (PPIX)-lipid micelles and liposomes in photodynamic therapy: S. Tachikawa, et al.; Bioorg. Med. Chem. 23, 7578 (2015), Application(s): Fluorescence microscopy analysis of HeLa cells, Abstract;
The Golgi apparatus is a functionally distinct Ca2+ store regulated by the PKA and Epac branches of the β1-adrenergic signaling pathway: Z. Yang, et al.; Sci. Signal. 8, ra101 (2015), Application(s): Confocal microscopy, Abstract; Full Text
Asp68His mutation in the A1 domain of human factor V causes impaired secretion and ineffective translocation: H.C. Liu, et al.; Haemophilia 20, e318 (2014), Abstract;
Synthesis of protoporphyrin–lipids and biological evaluation of micelles and liposomes: S. Tachikawa, et al.; Bioorg. Med. Chem. 22, 4745 (2014), Abstract;

General Literature References

Golgi-disturbing agents: A. Dinter & E.G. Berger; Histochem. Cell Biol. 109, 571 (1998), Abstract;
Fluorescent conjugates of brefeldin A selectively stain the endoplasmic reticulum and Golgi complex of living cells: Y. Deng, et al.; J. Histochem. Cytochem. 43, 907 (1995), Abstract;

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