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Product Details
| Cat # +Size | K120-50 |
|---|---|
| Size | 50 assays |
| Detection Method | Agarose gel with Ethidium bromide |
| Species Reactivity | Mammalian cell types |
| Applications | Efficiently detects fragmented DNA from apoptotic cells |
| Features & Benefits | • Simple one-step procedure; takes only 90 minutes • Fast and convenient • DNA fragmentation can be easily visualized by agarose gel electrophoreses. The new procedure increases recovery of small fragmented DNA, and therefore improves the sensitivity of the assay. |
| Kit Components | • TE Lysis Buffer • Enzyme A Solution • Enzyme B (Lyophilized) • Ammonium Acetate Solution • DNA Suspension Buffer |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells. BioVision’s Quick Apoptotic DNA Ladder Detection Kit provides an easy and sensitive means for detecting DNA fragmentation in apoptotic cells. Unlike other commercially available kits that require 1-2 days to perform the procedure, the new detection method requires less than 90 minutes to prepare DNA, with neither extraction nor using columns.
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What is the difference between K120 and K170?
Both K120 and K170 have similar protocols. However, in K170, there is a centrifugation step at the end of the protocol at 16,000 g, which will eliminate the genomic DNA from the samples. Additionally, K170 requires more cells and therefore there will be more genomic DNA whereas in K120, the number of cells is less and therefore, interference from genomic DNA will be less.
Can I do labeling with another antibody post Annexin-V labeling? If I permeabilize after fixation and do another antibody labeling then my annexin staining disappears. Does this have to do with the fluorophore or the permeabilization step?
Double labeling of annexin V with a cell surface marker has been widely used successfully. It is important to remember that Annexin V staining is calcium dependent,. Therefore all buffers used after Annexin V stainig step should contain 1.8-2mM Ca+2. Fluorescence can also fade overtime, so it's better to perform the subsequent labeling steps in dark.
Can the incubation with Annexin V be extended for 30 m in at RT?
The incubation time can be extended to 30 min. However, a longer incubation period can sometimes result in more dead cells, which can give false positive results. Generally the incubation time should be limited to 10 min. After incubation, you can put the cells on ice if you need to wait a while to do the next step.
I am interested in the kit for high throuphput screening. Can we use a plate reader to detect Annexin V?
Plate reader will not help you in differentiating apoptotic cells from dead cells.
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.
Cantaluppi V et al (2009) J. Am. Soc. Nephrol. 19: 1904-1918.
Papineni S et al. (2008) Mol. Pharmacol. 73: 553-565.
Sordet O et al (2008) J. Biol. Chem. 283: 23200 - 23208.
Wang L et al (2008) Cancer Res. 68: 4377-4383.
Kang SG et al. (2007) J. Biol. Chem. 282: 6090-6097.
For more citations of this product click here
