Annexin V-EGFP Apoptosis Kit

Detects Apoptosis in live cells within 10 min.
Catalog #: K104 | abID: ab14153

Product Details

abID ab14153
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
• EGFP is a bright and photo-stable green fluorescent protein and the kit can differentiate apoptosis vs necrosis when performing both annexin V-EGFP and PI staining
Kit Components • Annexin V-EGFP
• 1X Binding Buffer
• Propidium Iodide (PI)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


The Annexin V-EGFP Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid 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 an enhanced green fluorescent protein (EGFP) fusion of annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on • Live cells. Detection can be analyzed by flow cytometry or by fluorescence microscopy with a FITC filter. EGFP is brighter and more photo-stable than other fluorescent reagents. The kit can differentiate apoptosis vs necrosis when performing both annexin V-EGFP 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-EGFP Apoptosis Detection Kit could be adapted to a 96 well plate fluorescent plate reader format?
aIt 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, incubate them, 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.
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.
Will trypsinizing the cells removed the phosphatidylserine?
The trypsin used in this kit should in no way affect the PS on the cell membrane.
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.
Jingsi Zhang, et al., (2018) Jia-Jian-Di-Huang-Yin-Zi decoction exerts neuroprotective effects on dopaminergic neurons and their microenvironment, Scientific Reports, Jun. 2018, 29959371
Talwar et al., Inhibition of Caspases Protects Mice from Radiation-induced Oral Mucositis and Abolishes the Cleavage of RNA-binding Protein HuR. J. Biol. Chem.,  Feb 2014; 289: 3487 - 3500.
Hatton, O. et. al. Syk Activation of Phosphatidylinositol 3-Kinase/Akt Prevents HtrA2-dependent Loss of X-linked Inhibitor of Apoptosis Protein (XIAP) to Promote Survival of Epstein-Barr Virus+ (EBV+) B Cell Lymphomas. J. Biol. Chem., Oct 2011; 286: 37368 - 37378.
Talwar, S. et. al. Caspase-mediated Cleavage of RNA-binding Protein HuR Regulates c-Myc Protein Expression after Hypoxic Stress. J. Biol. Chem., Sep 2011; 286: 32333 - 32343.
Zhu, H. et. al. Kit-Shp2-Kit signaling acts to maintain a functional hematopoietic stem and progenitor cell pool. Blood, May 2011; 117: 5350 - 5361.
For more citations of this product click here