Annexin V-FITC Apoptosis Kit

Detects Apoptosis & Necrosis within 10 min.
Catalog #: K101 | abID: ab14085

Product Details

abID ab14085
Detection Method Flow cytometry (Ex = 488 nm; Em = 530 nm) and fluorescence microscopy
Species Reactivity Mammalian
Applications Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.
Features & Benefits • Simple one step staining procedure in 10 minutes
• Fast and convenient
• Kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining
Kit Components • Annexin V-FITC
• 1X Binding Buffer
• Propidium Iodide (PI)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Annexin V Apoptosis Detection Assay 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.

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The protocol in the datasheet for flow cytometry doesnt mention a wash step after Annexin/PI addition. can i confirm that this is correct?? I.e, 5min incubation then straight into flow cytometer.
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.
I am looking to investigate cell apoptosis using 96 well plates. I was wondering if your Annexin V-FITC Apoptosis Detection Kit could be adapted to a 96 well plate fluorescent plate reader format?
It is possible to adapt the assay to a plate reader format, however, the sensitivity can vary from sample to sample. 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 then analyze. The cell number can vary from cell to cell and/or treatment protocol selected. Please take measures to prevent overcrowding in the wells and also aspirate solutions gently since the apoptotic cells tend to detach quickly.
Can you prelabel the cells and then follow apoptosis using this kit?
Unfortunately, the product has not yet been tested in this manner. We are unsure whether it might work and needs significant optimization with known controls and timepoints to determine if the kit is performing correctly in these conditions.
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.
Can the kit work on bacteria or yeast cells?
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.
Will trypsinizing the cells removed the phosphatidylserine?
The trypsin used in this kit should in no way affect the PS on the cell membrane.
I see green color with my control cells (no Annexin-FITC, only PI)?
As for the visualisation of PI under the FITC channel, this is going to depend on the exitation and emission filters/laser you are using. PI can be excited to about 50% efficiency in the 488nm range. So if you have a broad pass, or long pass filter in your FITC set you will see the signal from the PI. (FITC Ex/Em is 495nm/519nm and Propidium iodide is 535nm/617nm).
Hongyan Wang, Role of JNK and ERK1/2 MAPK signaling pathway in testicular injury of rats induced by di-N-butyl-phthalate (DBP). Biol Res., Aug 2019;  31387634.
Diego G, Ammonia sensitive SLC4A11 mitochondrial uncoupling reduces glutamine induced oxidative stress. Redox Biol, June 2019; 31254733.
Taghizadehghalehjoughi, Vincristine combination with Ca+2 channel blocker increase antitumor effects. Mol Biol Rep., April 2019; 30903573.
Mariateresa Fulciniti, Non-overlapping Control of Transcriptome by Promoter- and Super-Enhancer-Associated Dependencies in Multiple Myeloma. Cell Rep, Dec 2018;  30590042.
Haixia Zhang, UHRF1 mediates cell migration and invasion of gastric cancer. Biosci Rep, Dec 2018;  30352833.
For more citations of this product click here