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Zeta life/Annexin V-FITC/PI apoptosis assay kit/An0020

价格
面议
货号:An0020
浏览量:127
品牌:Zeta life
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商品描述
Overview

Annexin V-FITC/PI apoptosis assay kit

Packing specification

Product number: An0020, An0050, An0100

Specifications: 20T, 50T, 100T

Storage conditions

Stored at 4oC, valid for one year

Product Description

The Annexin V-FITC Apoptosis assaykit detects apoptotic cells by flow cytometry. The annexins are a group of homologous proteins which bind phospholipids in the presence of calcium. Annexin V-FITC is a fluorescent probe which binds to phosphatidylserine in the presence of calcium. Apoptosis, or programmed cell death, is a mechanism of cells used to negatively select cells that are deleterious to the host. The cellular changes involved in the process include loss ofphospholipid asymmetry during the early stages ofapoptosis. In living cells, phosphatidylserine is transported to the inside of the lipid bilayer by the Mg-ATP dependent enzyme, aminophospholipidtranslocase. At the onset of apoptosis, phosphatidyl- serine, which is normally found on the internal part of the plasma membrane, becomes translocated to the external portion of the membrane. The phosphatidyl- serine becomes available to bind to the annexin V-FITC conjugate in the presence of calcium.

The procedure consists of the binding of annexin V- FITC to phosphatidylserine in the membrane of cells, which are beginning the apoptotic process, and the binding of propidium iodide to the cellular DNA in cells where the cell membrane has been totally compromised. Apoptosis may be either spontaneous orinduced by incubating the cells with staurosporine. The cells are incubated with annexin V-FITC and propidium iodide. After a 10 minute incubation period at room temperature the cells are analyzed by flow cytometry. Annexin V-FITC is detected as a green fluorescence and propidium iodide is detected as a red fluorescence.

Properties

Reagents:

Reagents

20 assays

50 assays

100 assays

Storage

Annexin V-FITC

product code:FI001

100 μl

250 μl

500 μl

4°C in the dark

Propidium Iodide, PI

product code:PI002

200 μl

500 μl

1000 μl

4°C in the dark

Binding Buffer ( 4)

product code:BU002

4 ml

10 ml

20 ml

4°C

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Annexin V is a product of human origin; handle as if capable of transmitting infectious agents.

Preparation Instructions

Allow all kit components to reach room temperature before use.

Prepare 1× Binding Buffer by diluting 1 ml of the 10× Binding Buffer with 9 ml of deionized water.

Dissolve the staurosporine in DMSO to a concentration of 100 μg/ml.

Procedure

This procedure describes the induction of apoptosis in the Jurkat cell line, followed by measurement of phosphatidylserine. Perform the experiment using aseptic technique.

1. Induce apoptosis in a 1 x 106cells/ml suspension of Jurkat cells by the addition of 1 μg/ml staurosporine.

2. Establish a control of non-induced Jurkat cells at 1 x 106cells/ml for a zero time data point.

3. Incubate both Jurkat cell cultures for 1–2 hours in a 37 °C, 5% CO2incubator.

4. Wash the cells twice with DPBS.

5. Resuspend the cells in 1× Binding Buffer at a concentration of ~1 x 106 cells/ml.

6. Add 500 μl of the apoptotic cell suspension to a plastic 12 × 75 mm test tube.

7. Add 500 μl of the non-induced cell suspension to a second plastic 12 × 75 mm test tube.

8. Add 5 μl of Annexin V FITC Conjugate and 10 μl of Propidium Iodide Solution to each cell suspension.

9. Incubate the tubes at room temperature for exactly 10 minutes and protect from light.

10. Determine the fluorescence of the cells immediately with a flow cytometer. Cells, which are early in the apoptotic process, will stain with the Annexin V FITC Conjugate alone. Live cells will show no staining by either the Propidium Iodide Solution or Annexin V FITC Conjugate. Necrotic cells will be stained by both the Propidium Iodide Solution and Annexin V FITC Conjugate.

Background
ZETA LIFE公司成立于1989年,曾为日本较大临床单克隆抗体生产商MBL Biotech公司提供13年的生产血清经验,也是目前血清OEM供应商之一。每批血清均有完成的血源证明文件、兽医师检验证明及品管测试报告。专人负责测试追踪、建档,能轻易帮您挑选合适的批号。该血清在登记100级的无菌操作室内,均采用全自动方式制造生产,具有良好的可再现性以及可追溯性。美国zeta life进口ZETA LIFE胎牛血清现货促销产品货号:Z7185FBS-500产品产地:美国产品规格:500mlZ7185FBS-500适用范围:适用于以下细胞的培养。干细胞:1、BMSC         大鼠骨髓间充质干细胞2、BMSC         犬骨髓间充质干细胞3、PGCs         鸡原始生殖干细胞