Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)

Overview

  • Product name
    Annexin V-FITC Apoptosis Staining / Detection Kit
    See all Annexin V kits
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Direct
  • Assay time
    0h 10m
  • Product overview

    Abcam's Annexin V-FITC Apoptosis Staining / Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane 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 high affinity for PS. The one-step staining procedure takes only 10 minutes. Detection can be analyzed by flow cytometry or by fluorescence microscopy. The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
    Visit our FAQs page for tips and troubleshooting.

  • Tested applications
    Suitable for: FM, Flow Cytmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab14085 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent dilution.

Images

  • HeLa cells were harvested with trypsinization together with floating non-viable cells.  Cells were washed with PBS and  suspended in sodium citrate buffer 20 minutes prior to analysis. HeLa cells were treated with Mitoxantrone (MX) and MX +Imatinib for 48 hours. The samples were then stained with Annexin V-FITC Apoptosis Staining/Detection kit (ab14085). A FACSCalibur flow cytometer was used for cell cycle analysis.

     

  • PC3 cells were seeded at 106 cells/ml and incubated overnight and then treated with CTA095 at various concentrations for 24hours.  Apoptosis was then analyzed using Annexin-V FITC apoptosis detection kit (ab14085).

  • Annexin V-FITC/ PI staining of AG06173 primary fibroblasts.
    105 cells were used for analysis. Resuspended cells were incubated with Annexin V-FITC for 15 min in the dark. Propidium iodide was used as a counterstain to discriminate necrotic/ dead cells from apoptotic cells. Left: negative control - AG6173 untreated cells. Right: positive control - AG6173 cells irradiated at 10 Gy.
    Image courtesy of S. Khoronenkova PhD, Gray Institute for Radiation Oncology and Biology, University of Oxford, Oxford, UK.

References

This product has been referenced in:
  • Jia P  et al. Gab3 overexpression in human glioma mediates Akt activation and tumor cell proliferation. PLoS One 12:e0173473 (2017). Read more (PubMed: 28291820) »
  • Wang C  et al. Dual inhibition of PCDH9 expression by miR-215-5p up-regulation in gliomas. Oncotarget 8:10287-10297 (2017). Functional Studies . Read more (PubMed: 28055966) »

See all 53 Publications for this product

Customer reviews and Q&As

Abreviews
We used the Annexin V-FITC apoptosis detection kit to determine minor levels of cell death (using both the Annexin V-FITC and PI) in mouse cortical collecting duct cellss (mCCDs). Using this kit, we were able to detect cell death in less than 5% of all cells using fluorescent microscopy. In the image provided, mCCD cells were incubated with serum free medium for 48h. The green label on the plasma membrane (Annexin V-FITC) and the absence of nuclear red (PI) staining indicates apoptosis rather than necrosis.

Discount code: ABTRIAL-EBLDA
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Abcam user community

Verified customer

Submitted Aug 07 2015

Typically washing is not needed before analysing in the flow cytometer. However, if you wish to fix your cells, this can be done after incubating with Annexin V in binding buffer. The cells can then be washed in PBS and fixed in 2% PFA for 15 mins. Read More

Miss Sarah Sabir

Excellent Excellent 5/5 (Ease of Use)
Abreviews
U2OS cells were either treated with Staurosporine (0.5 uM) for 6 hours or left untreated in 6cm dishes. Cells were resuspended using PBS-EDTA and collected in a 15ml tube along with original culture media. Cells were centifuged at 1100rpm for 5 minutes and then resuspended in binding buffer before adding AnnexinV-FITC and PI as in protocol. Cells were incubated for 5 minutes at RT and then analysed.
Additional controls (graphs not shown): no stain, PI only, Annexin V only
Figure legend: U2OS cells were either A. Untreated, or B. Treated with 0.5uM Staurosporine for 6 hours. Percentage of Live (Q3), early apoptotic (Q4) and late apoptotic (Q2) populations are indicated.
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Ms. Sarah Sabir

Verified customer

Submitted Nov 26 2013

Regarding ab14085 Annexin V-FITC Apoptosis Detection Kit , I can confirm this assay can indeed be adapted to a plate reader format as well. For this particular assay, take 10^5 trypsinized cells into the plate wells, resuspend in 100 µl binding b...

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Thank you for your inquiry.

I can confirm that ab14082 (5X Annexin V-FITC Apoptosis Detection Reagent) and ab14084 (10X Annexin V Binding Buffer) are the same reagents as in the kit ab14085(Annexin V-FITC Apoptosis Detection Kit).

P...

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Thank you for contacting us.

I have had the opportunity to discuss this with the originator. Since this product has not yet been tested in this manner we are unsure whether it might work. We cannot rule this out but feel that a number of exp...

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Thank you for your email.

I discussed with my colleagues from the Scientific Support Team about the use of these kits in yeast and we agreed that the kit that is likely to work is the In situ Direct DNA Fragmentation Assay Kit (ab66108) but no...

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Thank you for contacting us.

Unfortunately, these kits have been standardized for mammalian cells and the lab do not have the protocol suggestion of how to fit these to yeast cells, since they have never tried that.

Sorry I couldn't b...

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Thank you for your call today and for letting us know about the trouble with this kit.

As we discussed, I'll be back in touch once I have more information about the QC procedures from the production lab.

I am sending a free of charg...

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Thank you for your inquiry.

Yes, this kit should work with all mammalian cells.

I hope this information is helpful. Please contact us with any other questions.

1-10 of 21 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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