Annexin V-FITC/PI

  • Annexin V-FITC/PI Apoptosis Assay kit PI0020, PI0050, PI0100

Annexin V-FITC/PI Apoptosis Assay kit PI0020, PI0050, PI0100

  • Product number: PI0020, PI0050, PI0100

    Specifications: 20T, 50T, 100T



  • Manual Download

Annexin V-FITC/PI Apoptosis Assay kit

Packing specification

Product number: PI0020, PI0050, PI0100

Specifications: 20T, 50T, 100T

Storage conditions

Stored at 4ºC, valid for one year

Reagents:

Reagents

20 assays

50 assays

100 assays

Storage

Annexin V-FITC

product code:AF001

100 μl

250 μl

500 μl

4°C in the dark

Propidium Iodide, PI

product code:PP002

200 μl

500 μl

1000 μl

4°C in the dark

Binding Buffer ( 4´ )

product code:BB002

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 106 cells/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 106 cells/ml for a zero time data point.

3. Incubate both Jurkat cell cultures for 1–2 hours in a 37 °C, 5% CO2 incubator.

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.

It can only be used for scientific research. It is forbidden to use it for human, animal or other purposes.


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