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APOAF

Sigma-Aldrich

Annexin V-FITC Apoptosis Detection Kit

Synonym(s):

Annexin V-FITC, Apoptosis probe FITC

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About This Item

UNSPSC Code:
12352207
NACRES:
NA.84

usage

(20 tests)

Quality Level

300

packaging

pkg of 1 kit

technique(s)

flow cytometry: suitable

application(s)

cell analysis
detection

detection method

fluorometric

shipped in

wet ice

storage temp.

2-8°C

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).

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.

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.

Other Notes

Allow all components to reach room temperature before use.

Kit Components Only

Product No.
Description
  • APOAFA

related product

Product No.
Description
Pricing

Storage Class Code

10 - Combustible liquids

Regulatory Information

常规特殊物品

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
0000389724
0000378389
0000378388
0000159278
0000378389

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  1. 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.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. 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.

  4. 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).

  5. 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.

  6. 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.

  7. 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.

  8. 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. 

  9. 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

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Denise C Arruda et al.
The Journal of biological chemistry, 287(18), 14912-14922 (2012-02-16)
Complementarity-determining regions (CDRs) from monoclonal antibodies tested as synthetic peptides display anti-infective and antitumor activities, independent of the specificity of the native antibody. Previously, we have shown that the synthetic peptide C7H2, based on the heavy chain CDR 2 from
Monika Kallubai et al.
Journal of biomolecular structure & dynamics, 37(3), 623-640 (2018-01-30)
Our study focus on the biological importance of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules, the cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells
Ben-Zion Zaidman et al.
International journal of oncology, 31(4), 959-967 (2007-09-06)
Ganoderma lucidum (Curt.:Fr.) P. Karst, a medicinal fungus, has been widely used in Asian countries for centuries to prevent or treat a variety of diseases, including cancer. However, the mechanisms responsible for the effects of G. lucidum on cancer cells
Juliana Rizzo et al.
Eukaryotic cell, 13(6), 715-726 (2013-12-18)
Flippases are key regulators of membrane asymmetry and secretory mechanisms. Vesicular polysaccharide secretion is essential for the pathogenic mechanisms of Cryptococcus neoformans. On the basis of the observations that flippases are required for polysaccharide secretion in plants and the putative
Kalvin J Gregory et al.
PloS one, 5(10), e13428-e13428 (2010-10-27)
Vitamin D binding protein-macrophage activating factor (DBP-maf) is a potent inhibitor of tumor growth. Its activity, however, has been attributed to indirect mechanisms such as boosting the immune response by activating macrophages and inhibiting the blood vessel growth necessary for
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