enogene/Annexin V-EGFP凋亡检测试剂盒/100T/EBA12100
市场价：
¥4800.00
美元价：
2400.00
产品分类：
病毒纯化试剂盒
公司分类：
Virus_purification_kit
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
Annexin V-EGFP Apoptosis Detection Kit PDF
Catalog Number:
EBA12100Amount:
100TBrief Intro:
he Annexin V-EGFP Apoptosis Detection Kit is based on the separated observation of different time of apoptosis cells. The most phosphatidylserines (PS) in cell membrane phospholipids translocate from the inner face to the outer face in the early period of apoptosis. Once PSs are on the plasma membrane surface, it can be easily detected by staining with an enhanced green fluorescent protein (EGFP) fusion with annexin V, a protein that has strong natural affinity with PS. In addition, Propidium Iodide (PI) is a nucleic dye. It cannot be through cellular membrane directly, but it can pass membrane of cells in middle and late period of apoptosis and stain them red on nuclei. Detection can be analyzed by flow cytometry and fluorescence microscopy with a FITC filter. EGFP is brighter and much more photo-stable than other fluorescent reagents such as FITC, etc.KEY FEACTURE●Easy to perform: rapid and simple procedures to perform ●High Precision: detect the cells in the early period of apoptosis, accurately differentiate and compute the ratio of normal cells, apoptosis cells and necrotic cells.High Stabilization: EGFP has high signals, hard to quench.Price:
240$Experimental Methods:
Annexin V-EGFP Apoptosis Detection Kit PROTOCOLA. Incubation of suspension cells with Annexin V-EGFP 1. Induce apoptosis by a desired method. 2. Collect 1～5x105 suspension cells by centrifugation at 2000 rpm for 5 min, or collect adherent cells by using trypsin without EDTA. 3. Wash cells twice with PBS (centrifugation at 2000 rpm for 5 min) 4. Resuspend cells in 500 µl Binding Buffer. 5. Add 5µl of Annexin V-EGFP and 5µl of propidium iodide (PI), and mix, respectively.5. Incubate at room temperature for 5～15 min ,away from light . Proceed as follow as C or D depending on method of analysis.B. Incubation of adherent cells with Annexin V-EGFPFor adherent cells, there are two methods.a.Enzyme digest1.Collect 1～5x105 adherent cells by using trypsin without EDTA. After digested, cells were kept in culture medium with serum to prevent trypsin further digest. 2.The follow procedures are the same as Step A3～A5.b.Directly Observation 1.Culture cells on a coverslip, and induce apoptosis with proper inducer directly. Set negative control.2.Rinse samples twice with PBS3.Add 5µl Annexin V-EGFP and 5µl Propidium Iodide into 500µl Binding Buffer, mix.4.Add the mixed reagent above on the coverslip and make it uniform. 5.Incubate in wet and dark surrounding for 5min.C. Detection by Fluorescence Microscopy 1. Place the stained cells from Step A.5, or Step B.b.5 on a glass slide. Cover the cells with a coverslip. 2. Observe the cells under a fluorescence microscope using a dual filter for FITC and rhodamine. Cells which have been bound with Annexin V-EGFP will show green in the plasma membrane. Cells which have lost membrane integrity will show red (PI) throughout the nucleus and a halo of green staining (EGFP) on the cell surface (plasma membrane). D. Quantification by Flow Cytometry Analyze Annexin V-EGFP binding by flow cytometry (Ex =488 nm; Em =530nm) using FITC signal detector (usually FL1) and PI staining by the phycoerythrin emission signal detector (usually FL2). Fluorescence Balance: normal cells as the control, balance fluorescence to reduce spectrum overlapping and set the situation of orientation system.
品牌介绍
enogene的β-actin小鼠单克隆抗体（A02）PDF目录编号：E12-045量：100微克/ 100微升克隆编号：A02背景：β-肌动蛋白是已鉴定出的六种不同的肌动蛋白同工型之一。在各种物种和组织的细胞中发现的肌动蛋白分子在免疫学和物理特性方面趋于非常相似。因此，针对β-肌动蛋白的抗体可用作蛋白质印迹的上样对照。但是，应注意，β-肌动蛋白的水平在某些细胞中可能不稳定。例如，β-肌动蛋白在脂肪组织中的表达非常低，因此不应将β-肌动蛋白用作这些组织的负荷控制。抗体形式：pH 7.4、150mM NaCl，0.02％叠氮化钠和50％甘油的磷酸盐缓冲盐水（不含Mg2 +和Ca2 +）中的小鼠IgG1。储存/稳定性：-20℃/ 1年保存特异性/敏感性：抗体可以检测内源性β-肌动蛋白。反应性：H，R，M，Mk，Dg，Ch，Hm，Rb，Pg，Sh应用范围：WB：1：5,000-10,000 IHC：1：200 IF：1：200
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