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Annexin V-APC/Cyanine7 / 7-AAD Apoptosis Detection Kit[230]

  • Cat.No.:E-CK-A230

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Introduction

Elabscience® Annexin V-APC/Cyanine7 / 7-AAD Cell Apoptosis Detection kit is developed to identify apoptotic and necrotic cells.

Annexin V is a member of the annexin family, which binds to phosphatidylserine (PS) in a calcium-dependent manner. Annexin V-APC/Cyanine7, the APC/Cyanine7-conjugated format, binds specifically to the PS on the outer leaflet of apoptotic cell membrane and can be detected with flow cytometry or fluorescence microscopy.

7-Amino Actinomycin D (7-AAD) has a high DNA binding constant and is efficiently excluded by intact cells. It is useful for DNA analysis and dead cell discrimination during flow cytometric analysis. Due to the loss of integrity of membrane, 7-AAD can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using Annexin V and 7-AAD.

Components

Cat. Products 20 Assays 50 Assays 100 Assays 200 Assays Storage
E-CK-A129 Annexin V-APC/Cyanine7 Reagent[129] 100 μL 250 μL 500 μL 1 mL 2~8°C
E-CK-A151 Annexin V Binding Buffer (10 ×) 1.4 mL×2 5.5 mL 11 mL 11 mL×2 2~8°C
E-CK-A162 7-AAD Viability Staining Solution[162] 100 μL 250 μL 500 μL 1 mL
Manual One Copy

Jurkat cells were treated with 5 μM Camptothecin and detected with this kit.

Jurkat cells were cultured with (Left) or without (Right) 5 μM Camptothecin for 4 h. Annexin V-APC/Cyanine7 single-positive cells were early apoptotic cells, Annexin V-APC/Cyanine7 and 7-AAD double-positive cells were necrotic or late apoptotic cells, and 7-AAD single-positive cells were naked nuclei.

Instructions

Dilute Annexin V Binding Buffer (10 ×) with DI water to 1×Annexin V Binding working solution before use.
For example, take 1 mL Annexin V Binding Buffer (10 ×) [E-CK-A151] and add it to 9 mL DI water to get 10 mL Annexin V Binding working solution.

Storage

Store at 2~8°C for one year. Annexin V-APC/Cyanine7 and 7-AAD Staining Solution should be stored in the dark.  

Cautions

1.For maximal assay performance, this kit should be used within 12 months. Avoid freeze / thaw cycles.

2.For FCM analysis, please set untreated cells stained with both Annexin V-APC/Cyanine7 and 7-AAD as negative control. As for compensation controls, please use drug-treated cells stained with either Annexin V-APC/Cyanine7 or 7-AAD.

3.Annexin V-APC/Cyanine7 can be detected in APC/Cy7 channel while 7-AAD can be detected in PerCP/Cy5.5 channel.

4.Detect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis.

5.Avoid extended exposure of the samples to direct light to protect the fluorophores from quenching.

6.For your safety and health, please wear the lab coat and disposable gloves before the experiments.

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

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