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APOAF Sigma

Annexin V-FITC Apoptosis Detection Kit

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Features and Benefits

Detects apoptosis earlier in the process than DNA-based assays such as TUNEL.
• Rapid labeling of cells. Cell staining takes only 10 minutes.
• No cell fixation or processing required, reducing the detection time and allowing the cells to be used for further study.
• Propidium iodide secondary dye is included with the kit to differentiate apoptotic cells from viable and necrotic cells.

General description

Annexin V-FITC kit allows fluorescent detection of annexin V bound to apoptotic cells and quantitative determination by flow cytometry. The AnnexinV-FITC kit uses annexin V conjugated with fluorescein isothiocyante (FITC) to label phosphatidylserine sites on the membrane surface. The kit includes propidium iodide (PI) to label the cellular DNA in necrotic cells where the cell membrane has been totally compromised. This combination allows the differentiation among early apoptotic cells (annexin V positive, PI negative), necrotic cells (annexin V positive, PI positive), and viable cells (annexin V negative, PI negative).

Other Notes

Allow all components to reach room temperature before use.

Application

Annexin V-FITC Apoptosis Detection Kit was used:
• in staining of LNCaP prostate cancer cells for measuring the G. lucidum extracts activity during the treatment of prostate cancer.
• for tumor cell labelling to study the inhibitory activity of DBP-maf (Vitamin D binding protein-macrophage activating factor) on prostate tumor cells.
• for indirect measurement of flippase activity.

Price and Availability


Your Key to Success

All labs need water

Kit component only

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10× Binding buffer SDS    
Propidium Iodide solution SDS    

Standard component

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AnnexinV-FITC solution SDS A9210
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
When using the Annexin V-FITC Apoptosis Detection Kit, Product APOAF, can I use any buffer for resuspending my cells?
The binding buffer included in the kit needs to be used for resuspending cells.  The buffer contains calcium chloride at a final (1X) concentration of 2.5 mM which is necessary for the binding of annexin V to phosphatidylserine.
What wavelength do I use to detect Annexin V-FITC and Propidium Iodide when using Annexin V-FITC Apoptosis Detection Kit, Product APOAF?
Annexin V FITC will have a maximum emission of 528 nm. This can be measured in the standard FITC Channel on a flow cytometer (FL1).  Propidium Iodide has a maximum emission of 620 nm.  This is measured on the short red channel on a flow cytometer (FL2 or FL3).
Can I use Product APOAF, Annexin V-FITC Apoptosis Detection Kit, to differentiate cells that are dead due to necrosis or apoptosis?
When using a kinetic study (various time points), you can show the progression of the cells from viable (annexin V FITC negative, PI negative), to annexin FITC positive, PI negative (membrane flip) to annexin V FITC positive, PI positive (dead).  If there are cells that are double positive when starting, it is not possible to guarantee that the cell death occurred due to apoptosis with this assay.
When using Product APOAF, Annexin V-FITC Apoptosis Detection Kit, I have cells that are Annexin V FITC negative and PI positive.  What are these cells?
It is not possible to have cells that are PI positive without the cells also being positive for Annexin V binding.  Cells are PI positive because the membrane has been compromised.  If this is the case, Annexin V can also enter the cell and bind to the PS on the internal cell membrane. The gating on the histogram for the FITC channel should be changed so that all cells that are PI positive are also Annexin V positive.
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
Can Product APOAF, Annexin V-FITC Apoptosis Detection Kit be used on fixed cells?
No. Product APOAF, Annexin V-FITC Apoptosis Detection Kit must be performed on live cells in order to measure Apoptosis. The assay is based on the externization of phosphatidylserine from the inner cell membrane to the outer cell membrane.  If the membrane is preturbed due to fixation, non-speciifc staining of the inner cell membrane might occur
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Protocols & Articles

Articles

Apoptosis Detection Kits

Programmed cell death is a fundamental process important in development, as well as a principle mechanism of tumor suppression. Apoptosis is triggered in non-malignant cells as a protective mechanism...
Dalit Weinstein-Fischer, Ph.D., Supervisor of Research & Development; Rina Altman, M.Sc., Scientist of Research & Development; Dorit Zharhary, Ph.D., Director of Research & Development
Biowire Spring 2012, 22–24
Keywords: Antibiotics, Apoptosis, Cancer, Condensations, Flow cytometry, Growth factors, Ligands, Methods, Phosphorylations, Separation, Transcription

Peer-Reviewed Papers
15

References

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