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

Reviews (3)Q&A (19)References (212)

Overview

  • Product name

    Annexin V-FITC Apoptosis Staining / Detection Kit
    See all Annexin V/ANXA5 kits

  • Sample type

    Adherent cells, Suspension cells

  • Assay type

    Direct

  • Assay time

    0h 10m

  • Product overview

    Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane. Analysis is by flow cytometry or fluorescence microscopy.


    The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.


    The Annexin V-FITC reagent contained in the kit is also available as Annexin V-FITC reagent ab14082.

  • Notes

    Soon after initiating apoptosis, cells translocate membrane phosphatidylserine molecules from the inner face of the plasma membrane to the cell surface. Phosphatidylserine on the cell surface is detected by staining with a fluorescent conjugate of Annexin V, a protein that has a high affinity for phosphatidylserine.

    For more apoptosis assays, review the full set of Annexin V assays, or the apoptosis assay and apoptosis marker guide.

  • Platform

    Flow cytometer, Fluorescence microscope

Properties

Images

  • Apoptosis in Mouse Cortical Collecting Duct Cells
    Apoptosis in Mouse Cortical Collecting Duct CellsImage courtesy of an anonymous abreview

    Ab14085 was used to determine minor levels of apoptosis (using both the Annexin V-FITC and PI) in mouse cortical collecting duct cellss (mCCDs). 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. Fluorescent microsocpy ws used to analyse the cells.

  • Flow cytometery analysis of treated HeLa cells for 48 hours
    Flow cytometery analysis of treated HeLa cells for 48 hoursAlpay et al., PLos One, 9(19), Fig 5B. Doi:10.1371/journal.pone.0105526 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

     This is a modified version of the original image

  • Analysis of apoptosis in prostate cancer cells following treatment with CTA095
    Analysis of apoptosis in prostate cancer cells following treatment with CTA095Guo W et al., PLoS One, 8(8). Fig7b, doi: 10.1371/journal.pone.0070910 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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).

    This is a modified version of the original image

  • Functional Studies - Annexin V-FITC Apoptosis Detection Kit (ab14085)
    Functional Studies - 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 (212)

Publishing research using ab14085? Please let us know so that we can cite the reference in this datasheet.

ab14085 has been referenced in 212 publications.

  • Cao Y  et al. Long Noncoding RNA UCA1 Regulates PRL-3 Expression by Sponging MicroRNA-495 to Promote the Progression of Gastric Cancer. Mol Ther Nucleic Acids 19:853-864 (2020). PubMed: 31982772
  • Li S  et al. Deficiency in Dipeptidyl Peptidase-4 Promotes Chemoresistance through the CXCL12/CXCR4/mTOR/TGFß Signaling Pathway in Breast Cancer Cells. Int J Mol Sci 21:N/A (2020). PubMed: 31991851
  • Yang C  et al. Silencing of microRNA-517a induces oxidative stress injury in melanoma cells via inactivation of the JNK signaling pathway by upregulating CDKN1C. Cancer Cell Int 20:32 (2020). PubMed: 32015692
  • Elias-Kirma S  et al. In situ-Like Aerosol Inhalation Exposure for Cytotoxicity Assessment Using Airway-on-Chips Platforms. Front Bioeng Biotechnol 8:91 (2020). PubMed: 32154228
  • Venit T  et al. Nuclear myosin 1 activates p21 gene transcription in response to DNA damage through a chromatin-based mechanism. Commun Biol 3:115 (2020). PubMed: 32161327
View all Publications for this product

Customer reviews and Q&As

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1-10 of 22 Abreviews or Q&A

Flow cytometery analysis of treated multiple myeloma cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Multiple myeloma cells were harvested by centrifugation after collecting the trypsinized cellls together with floating non-viable cells. Cells were washed with PBS and re-suspended in sodium citrate buffer 20 minutes prior to analysis. The cells were then treated with Mitoxantrone (MX) and MX +Imatinib for 24 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.

Abcam user community

Verified customer

Submitted Jun 25 2018

Apoptosis in mouse cortical collecting duct cells

 Good Excellent 5/5 (Ease of Use)
Abreviews
abreview image
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

Abcam user community

Verified customer

Submitted Aug 07 2015

Question

the protocol booklet for the Annexin v fit-c kit for flow cytometry doesnt mention a wash phase after antibody/PI addition. can i confirm that this is correct?? I.e, 5min incubation then straight into flow cytometer? to fix following this, is 4% PFA recommended?

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Answer

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.

Once your cells have been re-suspended in PBS, you can also add a preservative/stabiliser if you are planning on keeping your samples for a long time. For example, 1 mg/ml BSA or 1% sodium azide.

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Miss Sarah Sabir

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
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.

Ms. Sarah Sabir

Verified customer

Submitted Nov 26 2013

Question

ab14085

I am looking to investigate cell apoptosis using 96 well plates. I was wondering if your Annexin V-FITC Apoptosis Detection Kit could be applied to such a format. I notice the protocol only includes flow cytometry and microscopy, but I am thinking it could be adapted to a 96 well plate and fluorescent plate reader format.

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Answer

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 buffer, add 1µ Annexin V and PI respectively, incubate them in the dyes, give a gentle wash and they are ready to be analyzed.

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Question

Dear All,
a customer of ours bought ab14085 (Annexin V apoptosis detection kit), but he needs to buy Annexin V-Fitc and Annexin V binding buffer separately.
Can you please confirm that the following codes are the same included within the kit ab14085:


Annexin V-Fitc #ab14082

Annexin V binding buffer #ab14084

Thanks in advance and kind regards

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Answer

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).

Please note that ab14082 (5X Annexin V-FITC Apoptosis Detection Reagent) and ab14084 (10X Annexin V Binding Buffer) are more concentrated and need need to be diluted to 1x before using them.

I hope this information is helpful and wish you and your customer a good week.

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Question

Customer kindly contacted us to inquire is they might use this product in the following manner. Add the kit components to live cells followed by treatment of the cell with a chemical to induce apoptosis and then read. They were further  wondering how long the dye might remain viable.

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Answer

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 experiments would need to be performed with known controls and timepoints to determine if the kit is performing correctly in these conditions. I am sorry that we have been unable to provide a better answer to you at this time regarding this product and hope that you continue to contact us regarding any Abcam product.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.



Use our products? Submit an Abreview. Earn rewards!

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Question

Can be it offer a testing discount for each kits If the user would like to test the kits, Annexin V-FITC Apoptosis Detection Kit (ab14085) and In situ Direct DNA Fragmentation Assay Kit (ab66108), on yeast cells by IF?

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Answer

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 not the Annexin V-FITC Apoptosis Detection Kit (ab14085).

I have therefore created the following testing discount code for ab66108 to be tested in yeast:

DISCOUNT CODE: ******
Expiration date: DD MM YYYY

In order to link the discount code to the Abreview that will be submitted, could you please send me the email address as well as the name of the institute of your customer?

This code will give your customer 1 free kit or value off of order before the expiration date. To redeem this offer, your customer must submit an Abreview for yeast and include this code in the “Additional Comments” section so we know the Abreview is for this promotion.
The submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed on a future order.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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Question

Are the kits, Annexin V-FITC Apoptosis Detection Kit (ab14085) and In situ Direct DNA Fragmentation Assay Kit (ab66108), suitable for immunofluorosence application on yeasts? Do you have a protocol suggestion specific for yeasts applications?

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Answer

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 be more helpful. I hope this information isneverthelesshelpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Kits have been successfully used in the past but two recently-ordered kits have shown no signal.

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Answer

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 charge kit on the order 1119340 which will arrive tomorrow.

I look forward to hearing from you. If there is anything else that we can do for you, please let me know and I'll be happy to help.

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1-10 of 22 Abreviews or Q&A

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