Annexin V-APC/PI Apoptosis Kit
中文名称:Annexin V-APC / PI细胞凋亡检测试剂盒
Price:
- 应用: 细胞凋亡-Annexin V
检测原理 | Annexin V是一种钙离子依赖性磷脂结合蛋白,与磷脂酰丝氨酸PS有高度亲和力,可通过细胞外侧暴露的PS与凋亡早期细胞胞膜结合,将Annexin V标记荧光染料,利用流式细胞仪检测细胞凋亡。
细胞核染料(PI,DAPI,7-AAD)可与双链DNA特异性结合,并产生强烈的荧光,正常情况下无法透过细胞膜。由于凋亡晚期或坏死细胞膜丧失完整性,细胞核染料(DAPI、PI、7-AAD)可进入细胞内对DNA进行染色,与Annexin V搭配使用,可区分处于不同凋亡时期的细胞。 |
检测方法 | 荧光;流式 |
样本类型 | 细胞 |
检测时间 | 40min |
检测仪器 | 流式细胞仪 |
荧光 | APC |
Ex/Em | 650/660 |
Channel set | APC |
自备试剂 | PBS(E-BC-R187), 去离子水 |
保存温度 | 该产品可在 2~8°C避光条件下保存12个月。 |
有效期 | 12个月 |
运输条件 | 冰袋 |
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E-CK-A117 | Annexin V-APC Reagent | 500Tests , 200Tests , 100Tests , 50Tests |
E-CK-A151 | Annexin V Binding Buffer (10×) | 50mL , 10mL |
E-CK-A218 | Annexin V-APC/7-AAD Apoptosis Kit | 200Assays , 100Assays , 50Assays , 20Assays |
E-CK-A258 | Annexin V-APC/DAPI Apoptosis Kit | 200Assays , 100Assays , 50Assays , 20Assays |
E-CK-A301 | Mitochondrial Membrane Potential Assay Kit (with JC-1) | 100Assays , 50Assays , 20Assays |
E-CK-A320 | One-step TUNEL In Situ Apoptosis Kit (Green, FITC) | 100Assays , 50Assays , 20Assays |
Q1:Can PBS be used to replace Binding Buffer in Annexin V assay?
No, because Annexin V binds to phosphatidylserine (PS) dependent on Ca2+, which is not present in common PBS solutions, while Binding buffers contain Ca2+ to facilitate Annexin V binding to PS. Without Binding Buffer, Annexin V binding capacity will be reduced, resulting in significantly reduced positive signal or even no positive signal.
Q2:Can Annexin V kit be used on adherent cells?
Annexin V kit can be used on adherent cells, with the following caveat: 1. In late stage of apoptosis, cells may not able wo maintain adhesion, so floating cells should be collected for the test. 2. The dissociation of adherent cells should be handled gently to avoid cell damage caused by human operation. 3. Do not digest for too long with trypsin, avoid the usage of the trypsin contain EDTA. If EDTA-containing trypsin must be used, wash the cells to remove EDTA as much as possible.
Q3:Is it able to detect blood samples with Annexin V assay kit?
It is not recommended to do so. Platelet and some other immune cells in blood may have exposed PS in normal state that will affect the test results.
Q4:How to set the gate for rathippocampal neuron apoptosis assay?
Annexin V assay is normally used for cell line samples, thus only debris need to be excluded by gating. Since there aremultiple types of cells in tissue samples, it depends on the purpose of experiment, whether it means to detect apoptosis of the whole tissue sample or some specific cells. Cells can be gated by FSC/SSC or some other biomarkers. Besides, operation of tissue sample preparation may also effect the result, Inappropriate operation may also result in false positives due to sample handling.
Q5:Annexin V Kit can be used to detect plant samples?
There is no such experience as verification yet. It is recommanded to take a pre-test if necessary.
Q6:Is Annexin V Kit able to detect cryopreserved cells directly?
It is not recommended to do so. Cryopreservation may lead to apoptosis and necrosis, resulting in inaccurate experimental results.