Annexin V-PE-Cy5 Apoptosis Detection Kit

Detects Apoptosis in • Live cells within 10 min.
SKU: K129

Availability: In stock

SKU-Size Size Price Qty
K129-25 25 assays
$175.00
K129-100 100 assays
$365.00
K129-400 400 assays
$865.00

Details

The Annexin V-PE-Cy5 Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can be easily detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on • Live cells, without the need for fixation. Results can be analyzed by flow cytometry or fluorescence microscopy.

Product Details

Detection Method Flow cytometry (Ex = 488 nm; Em = 670 nm) and fluorescence microscopy
Species Reactivity Mammalian
Applications Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.
Features & Benefits • Simple one-step procedure; takes only 10 minutes
• Fast and convenient, no need of fixation of cells
Kit Components • Annexin V-PE-Cy5
• 1X Binding Buffer
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.
Can I do labeling with another antibody post Annexin-V labeling? If I permeabilize after fixation and do another antibody labeling then my annexin staining disappears. Does this have to do with the fluorophore or the permeabilization step?
Double labeling of annexin V with a cell surface marker has been widely used successfully. It is important to remember that Annexin V staining is calcium dependent,. Therefore all buffers used after Annexin V stainig step should contain 1.8-2mM Ca+2. Fluorescence can also fade overtime, so it's better to perform the subsequent labeling steps in dark.
Can the incubation with Annexin V be extended for 30 m in at RT?
The incubation time can be extended to 30 min. However, a longer incubation period can sometimes result in more dead cells, which can give false positive results. Generally the incubation time should be limited to 10 min. After incubation, you can put the cells on ice if you need to wait a while to do the next step.
I am interested in the kit for high throuphput screening. Can we use a plate reader to detect Annexin V?
Plate reader will not help you in differentiating apoptotic cells from dead cells.
For this kit, where was the Annexin-V obtained from? Is it recombinant? Or purified? And from what species?
The annexin V is a recombinant protein expressed from E. coli. The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
The expiry date is typically 1 year from the date of shipment
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Kakhlon O et al (2008) Blood; 112: 5219 - 5227.
Yue Z et al (2008) J. Cell Biol.; 183: 279 - 296.
Reininger, A.J., et al. (2006) Blood 10.1182/blood-2005-02-0618.
For more citations of this product click here
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