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Product Details
| Detection Method | Flow cytometry (Ex = 649 nm; Em = 670 nm) and fluorescence microscopy |
|---|---|
| Species Reactivity | Mammalian |
| Applications | N/A |
| Features & Benefits | • Simple one step staining procedure in 10 minutes • Fast and convenient • Annexin V-Cy5 will show bright red-blue staining on the plasma membrane |
| Kit Components | • Annexin V-Cy5 • 1X Binding Buffer |
| Storage Conditions | 4°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Annexin V Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidyl-serine (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.
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I am looking to investigate cell apoptosis using 96 well plates. I was wondering if your Annexin V-Cy5 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, 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.
Sakuma, Brittany N. et al. (2016) Quantitative evaluation of malignant gliomas damage induced by photoactivation of IR700 dye, Sci Technol Adv Mater. 2016 Aug 22;17(1):473-482.
Wang, Rui et al. (2016) Association of serum pentraxin 3 concentrations with diabetic nephropathy J Investig Med. 2016 Aug;64(6):1124-7.
Fujii et al., TMEM16F is required for phosphatidylserine exposure and microparticle release in activated mouse platelets. PNAS, Oct 2015; 112: 12800 - 12805.
Samejima et al., Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death: A CELL SUICIDE MODULE. J. Biol. Chem., Nov 2014; 289: 31617 - 31623.
Dyugovskaya et al., The development of giant phagocytes in long-term neutrophil cultures. J. Leukoc. Biol.,Oct 2014; 96: 511 - 521.
For more citations of this product click here
