Rhod-3 Calcium Imaging Kit
Rhod-3 Calcium Imaging Kit
Citations & References (19)
Invitrogen™

Rhod-3 Calcium Imaging Kit

The novel Rhod-3 Calcium Imaging Kit is designed for live-cell imaging of cytosolic calcium signaling in combination with green-fluorescent dyesRead more
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Catalog NumberQuantity
R101451 Kit
Catalog number R10145
Price (EUR)
568,00
Each
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Quantity:
1 Kit
Price (EUR)
568,00
Each
Add to cart
The novel Rhod-3 Calcium Imaging Kit is designed for live-cell imaging of cytosolic calcium signaling in combination with green-fluorescent dyes or proteins.The red-fluorescent Rhod-3 AM dye supplied in the Rhod-3 Calcium Imaging Kit offers significant improvements over existing Ca2+-sensing dyes. The uptake into organelles has been reduced, which translates into better cytosolic localization. Also, Rhod-3 AM exhibits a large increase in fluorescence emission upon binding to calcium, making it more sensitive than other red-fluorescent calcium dyes.

Learn more about ion indicators including calcium, potassium, pH, and membrane potential indicators ›

  • Compared to existing red calcium dyes, Rhod-3 AM exhibits a large increase (>2.5 fold) in fluorescence upon binding Ca2+ and very low fluorescence at rest (without Ca2+ binding).

  • Narrow emission enables simultaneous detection with GFP or other dyes.

  • Enhanced cytosolic localization for better detection of cytosolic transients

    For easy cell loading, the Rhod-3 Calcium Imaging Kit contains PowerLoad™ Concentrate. Due to the unique nature of the PowerLoad™ solution, it can be used in the presence of complete culture media, thus reducing the negative effects of replacing media or loading in serum-free media.

    A proprietary, water-soluble form of probenecid (which is commonly used to inhibit cellular transport and thus reduces the baseline signal) is also supplied with the Rhod-3 Calcium Imaging Kit.

    Calcium indicators are important tools in signal transduction and cell-based pharmacological screening. In cells and tissues with blue or green autofluorescence, long-wavelength (i.e., red-shifted) calcium indicators provide a means to bypass this overlapping autofluorescence. Additionally, red-shifted calcium dyes are well suited for calcium imaging experiments multiplexed with green-fluorescent protein (GFP) or other green fluorescent dyes.

    The Rhod-3 AM dye (included in the Rhod-3 Calcium Imaging Kit) is an improved red-shifted rhodamine-based calcium dye with an attached AM ester group. Once inside the cell, intracellular esterases cleave the AM group, trapping the dye in the cell and rendering the dye fluorescent. As calcium binds, the fluorescence intensity of the dye is enhanced. The spectral properties of Rhod-3 AM (ex/em = ∼560/600 nm) and strong fluorescence upon calcium binding provide an excellent signal window and make it our brightest red calcium dye. Additionally, Rhod-3 AM displays a more uniform cytosolic distribution and improved signal compared to existing red calcium dyes such as Rhod-2.

    • For Research Use Only. Not for use in diagnostic procedures.
    Specifications
    Detection MethodFluorescence
    Dye TypeFluorescent Dye-Based
    FormLyophilized
    ModificationUn-Modified, Un-Modified
    Quantity1 Kit
    Shipping ConditionRoom Temperature
    Sub Cellular LocalizationCytoplasm
    ColorRed
    EmissionVisible
    For Use With (Application)Calcium Assay
    For Use With (Equipment)Fluorescence Microscope
    Product LineMolecular Probes
    Product TypeDye
    Unit SizeEach
    Contents & Storage
    The kit contains sufficient reagents for staining 50 coverslips based upon the protocol provided. Store kit at -20°C, desiccated and protected from light. When stored as directed, this kit is stable for 1 year.

    Frequently asked questions (FAQs)

    When using Calcium Crimson, AM, as an indicator of intracellular calcium flux, I am not getting a good degree of change. The cells also have GFP. What can I do?

    This is a known drawback of Calcium Crimson, AM (as well as Calcium Orange, AM and Fura Red, AM, which are also in the same emission range). You can try increasing the concentration and washing out of any unlabeled dye from the media, to try to get better signal-to-background. If that fails, we recommend using Rhod-3 AM instead, which has a much better change in signal in that wavelength.

    Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

    Why don't I see a significant change in signal for my live-cell fluorescent indicator dye?

    Regardless of the type of live-cell indicator dye (e.g., calcium indicators, pH indicator, metal ion indicators), make sure there is no serum during the loading step, which can prematurely cleave dyes with AM esters and bind dyes non-specifically. Always optimize the dye concentration and staining time with a positive control before you run your test samples, to give the best signal-to-background. Always run a positive control with a buffer containing free ions of known concentration and an ionophore to open pores to those ions (for instance, for calcium indicators like Fluo-4 AM, this would include a buffer with added calcium combined with calcimycin, or for pH indicators, buffers of different pHs combined with nigericin). Reactive oxygen indicators, such as CellROX Green or H2DCFDA would require a cellular reactive oxygen species (ROS) stimulant as a positive control, such as menadione. Finally, make sure your imaging system has a sensitive detector. Plate readers, for instance, have much lower detector efficiency over background, compared to microscopy or flow cytometry.

    Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

    Citations & References (19)

    Citations & References
    Abstract
    Modulation by Syk of Bcl-2, calcium and the calpain-calpastatin proteolytic system in human breast cancer cells.
    Authors:Fei B, Yu S, Geahlen RL,
    Journal:
    PubMed ID:23684705
    'Syk is a 72kDa non-receptor tyrosine kinase that is best characterized in hematopoietic cells. While Syk is pro-tumorigenic in some cancer cell types, it also has been reported as a negative regulator of metastatic cell growth in others. An examination of the RelA (p65) subunit of NF-?B expressed in MCF7 ... More
    Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors.
    Authors:Ieda M, Fu JD, Delgado-Olguin P, Vedantham V, Hayashi Y, Bruneau BG, Srivastava D,
    Journal:Cell
    PubMed ID:20691899
    'The reprogramming of fibroblasts to induced pluripotent stem cells (iPSCs) raises the possibility that a somatic cell could be reprogrammed to an alternative differentiated fate without first becoming a stem/progenitor cell. A large pool of fibroblasts exists in the postnatal heart, yet no single ' ... More
    Induction of human cardiomyocyte-like cells from fibroblasts by defined factors.
    Authors:Wada R, Muraoka N, Inagawa K, Yamakawa H, Miyamoto K, Sadahiro T, Umei T, Kaneda R, Suzuki T, Kamiya K, Tohyama S, Yuasa S, Kokaji K, Aeba R, Yozu R, Yamagishi H, Kitamura T, Fukuda K, Ieda M,
    Journal:
    PubMed ID:23861494
    'Heart disease remains a leading cause of death worldwide. Owing to the limited regenerative capacity of heart tissue, cardiac regenerative therapy has emerged as an attractive approach. Direct reprogramming of human cardiac fibroblasts (HCFs) into cardiomyocytes may hold great potential for this purpose. We reported previously that induced cardiomyocyte-like cells ... More
    MKP3 eliminates depolarization-dependent neurotransmitter release through downregulation of L-type calcium channel Cav1.2 expression.
    Authors:Mortensen OV,
    Journal:Cell Calcium
    PubMed ID:23337371
    'Release of neurotransmitters is a fundamental and regulated process that is essential for normal brain functioning. Regulation of this process is potentially important for any neuronal process, and disruption of the release process may contribute to the pathophysiology associated with psychiatric diseases. In this work it is shown that expression ... More
    Neprilysin deficiency protects against fat-induced insulin secretory dysfunction by maintaining calcium influx.
    Authors:Zraika S, Koh DS, Barrow BM, Lu B, Kahn SE, Andrikopoulos S,
    Journal:Diabetes
    PubMed ID:23328128
    'Neprilysin contributes to free fatty acid (FFA)-induced cellular dysfunction in nonislet tissues in type 2 diabetes. Here, we show for the first time that with prolonged FFA exposure, islet neprilysin is upregulated and this is associated with reduced insulin pre-mRNA and ATP levels, oxidative/nitrative stress, impaired potassium and calcium channel ... More
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