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Index / Biovis / Annexin V_Cy5 Reagent / Product Detail : 1013-1000 Annexin V_Cy5 Reagent
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#1013-1000 Annexin V_Cy5 Reagent

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  Price : 1152   EUR
1307   USD
893   GBP
4840   Zloty
154161   JPY
8885   NOK
9520   SEK
1303   CHF

Product name : Annexin V_Cy5 Reagent

Catalog number : 1013-1000

Quantity: 1000 assays

Availability: Yes

Supplier name : Biovis

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More Details about

STORAGE CONDITIONS: Store at 4°C. Do not freeze.
SHELF LIFE: 1 year under proper storage conditions


DESCRIPTION:
Annexin V-Cy5 is a bright fluorescent reagent for detecting of the early stages of apoptosis. During apoptosis, phosphatidylserine (PS) is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for PS and therefore serves as a probe for detecting apoptosis. Cy5 fluorescent dye produces an intense signal in the far-red region of the spectrum and therefore it is very useful for multiple labeling of cells with green and red colored fluorescent probes. Cy5 yields fluorescence with a max emission of 670 nm.


ASSAY PROTOCOL:
A. Incubation of cells with Annexin V-Cy5:
1. Induce apoptosis by desired methods.
2. Collect 1 x 105 cells by centrifugation.
3. Resuspend cells in 500 μl of 1X Annexin V Binding Buffer (Cat.#1035-100).
4. Add 1 μl of Annexin V-Cy5.
5. Incubate at room temperature for 5 min in the dark.
Proceed to B or C below depending on method of analysis.
B. Quantification by Flow Cytometry:
Analyze cells by flow cytometry (Ex = 649 nm; Em = 670 nm) using Helium-Neon Laser. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-Cy5 (A.3-5).
C. Detection by Fluorescence Microscopy:
1. Place the cell suspension from Step A.5 on a glass slide, and cover with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on a glass slide and visualize cells. The cells can also be washed with 1X Annexin V Binding Buffer and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-Cy5 before fixation because any cell membrane disruption can cause nonspecific binding of annexin V to PS on the inner surface of the cell membrane.)
2. Observe the cells under a fluorescence microscope using Cy5 filter or a FITC/Cy3/Cy5 triple band filter set (Chroma Technology) if you perform triple labeling with these dyes, or detect cells using CCD camera.
Cells that have bound Annexin V-Cy5 will show bright red-blue staining on the plasma membrane.


FOR RESEARCH USE ONLY! Not to be used in humans!

 

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