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主营:微管蛋白、肌动蛋白、小G蛋白、GAPs、GEFs
℡ 4000-520-616
℡ 4000-520-616
Cytoskeleton/SPY555-DNA/100 slides SPY555-DNA/CY-SC201
产品编号:CY-SC201
市  场 价:¥6500.00
场      地:美国(厂家直采)
产品分类: 蛋白类>多肽>多肽合成>
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美  元  价:$325.00
品      牌: Cytoskeleton
公      司:Cytoskeleton
公司分类:
Cytoskeleton/SPY555-DNA/100 slides SPY555-DNA/CY-SC201
商品介绍
Details

SPY555-DNA is a bright & non toxic live cell nuclear stain based on our SPYTM dyes series. Its optimized structure allows quick labeling of DNA in live & fixed cells with high specificity and very low background. SPY555-DNA stains the nuclei of live or fixed cells without the need for genetic manipulation or overexpression of fluorescent proteins. Its absorbance and emission are similar to tetramethyl-rhodamine (TMR). SPY555-DNA enables multicolor imaging with SPY505, SPY595, SPY650, SPY700, SIR and GFP. SPY555-DNA can be imaged with standard TMR or Cy3 filtersets. It can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue. Contains 1 vial of SPY555-DNA (lyophilized).

Absorbance maximum λabs

555 nm

Fluorescence maximum λfl

580 nm

Works on fixed cells?

yes, PFA and methanol

Probe quantity

100 stainings*

Fluorescence lifetime

2.4 ns

STED depletion wavelength

660 or 775 nm

Shipping

room temperature

Storage

-20°C

555-DNA

Comparison of SPY-labeled HeLa cells imaged with confocal vs. STED microscopy. Imaged with 93X objective and provided courtesy of Spirochrome.

About

For product Datasheets and MSDSs please click on the PDF links below.

Spirochrome Technical Tips and Ex/Em spectra in graphical form (PDF)

Ex/Em spectra (raw data in Excel format)

Citations

Featured Papers

“Fluorogenic probes for live-cell imaging of the cytoskeleton”; G. Lukinavičius, L.Reymond, E. D’Este, A. Masharina, F. Göttfert, H. Ta, A. Güther, M. Fournier, S. Rizzo, H. Waldmann, C. Blaukopf, C. Sommer, D. W. Gerlich, H.-D. Arndt, S. W. Hell & K. Johnsson; Nature Methods 11, 731–733

Faqs

Q1. What is STED microscopy and how does it work?

A1. STED microscopy stands for Stimulated Emission Depletion microscopy.  It is one type of super resolution microscopy which allows the capture of images with a higher resolution than conventional light microscopy which is constrained by diffraction of light.  STED uses 2 laser pulses, one is the excitation pulse which excites the fluorophore, causing it to fluoresce.  The second pulse, referred to as the STED pulse, de-excites the fluorophore via stimulated emission in an area surrounding a central focal spot that is not de-excited and thus continues to fluoresce.  This is accomplished by focusing the STED pulse into a ring shape, a so-called donut, where the center focal spot is devoid of the STED laser pulse, conferring high resolution to the fluorescent area (Fig. 1; see Ref. 1 for more details on STED microscopy).

fig1-faq

Figure 1. STED microscopic image of microtubules labeled with SiR-tubulin in human primary dermal fibroblasts.

Q2. Why is the SPY actin (or tubulin/DNA) probe good for STED microscopy?

A2. STED microscopy offers the ability to study cellular details on a nanometermolar scale in vivo.  To take advantage of this super resolution microscopy, one must be able to select with high specificity the area to be examined using fluorescent probes.  In addition, the fluorescent probes must be bright, photostable, exhibit no or little phototoxicity, be excited and emit in the far red spectrum.  In addition, if the probe is to be used for live cell imaging (thus avoiding fixation artifacts that occur when cells are fixed), high cell permeability is necessary.  The SPY actin and tubulin probes fulfill all of these requirements.  In short, the combination of STED and SiR probes allows for unparalleled fluorescent visualization of subcellular actin and tubulin/microtubule structures and their physical characterization in living cells, (see Fig. 2 and Ref. 2). 

Neuron_actin_1
Neuron_actin_closup2_1

Figure 2. STED images of cultured rat hippocampal neurons stained with SiR-actin. Bottom image is a close-up view of part of the top image to clearly visualize actin rings (stripes) with 180 nm periodicity. Courtesy Of Elisa D"Este, MPI Biophysical Chemistry, Göttingen.

Q3: Are the SPY probes stable at room temperature?

A3: Yes, the probes are stable at room temperature for a few days.  However, it strongly depends on the probe and the solvent.  Thus, it is recommended to store all of the probes or solutions at –20°C.

 

Q4: Are SPY-actin, SPY-DNA and SPY-tubulin toxic to cells?

A4: Yes, above a certain threshold both probes show some effect on cell proliferation and altered actin or microtubule dynamics.  However, the probes are orders of magnitude less toxic than their parent drug.  In HeLa cells, neither actin nor microtubule dynamics were altered at concentrations below 100 nM.  At this concentration, SPY probes efficiently label microtubules and F-actin, allowing for the capture of high signal to noise images.

References

1. Hell S.W. and Wichmann J. 1994. Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19, 780-782.

2. D’Este E. et al. 2015. STED nanoscopy reveals the ubiquity of subcortical cytoskeleton periodicity in living neurons. Cell Rep. 10, 1246-1251.

3. Lukinavicius G. et al. 2013. A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins. Nat. Chem. 5, 132-139.

4. Lukinavicius G. et al. 2014. Fluorogenic probes for live-cell imaging of the cytoskeleton.Nature Methods. 11, 731-733.

品牌介绍

Cytoskeleton公司成立于1993年,专注于生物化学和细胞过程研究中的纯化蛋白和便捷试剂盒开发与生产。公司提供药物筛选、信号转导、细胞骨架研究相关的系列试剂盒和产品,尤其以细胞骨架相关研究见长,既能满足于样品较少的科学研究,也可以用于小规模筛选研究和高通量大规模筛选研究。此外,公司还提供微管蛋白,肌动蛋白,小G蛋白,GAPs,GEFs等现有产品的药物筛选服务。

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