Apoptosis tools

- Prices without shipping charges & VAT -

single tool

tests

order No. / data sheet

price €

Annexin-V-FITC

100

AnxF100

100 €

Annexin-V-PE

100

AnxP100

100 €

Annexin-V-APC

100

AnxA100

100 €

Annexin-V-Biotin

100

AnxB100

100 €

Propidium iodide

100

PI100

50 €

Annexin-V binding buffer (10x)

10 ml

AnxBuff

30 €

Kit

tests

order No. / data sheet

price €

Annexin-V-FITC / Propidium iodide

100

AnxF100PI

160 €

Annexin-V-APC / Propidium iodide

100

AnxA100PI

160 €

Annexin-V-Biotin / Propidium iodide

100

AnxB100PI

160 €

IMG_2952

Ready-to-use

Annexin-V Apoptosis Detection Kit

Fast analysis of apoptosis in single cell suspensions

Background

MabTag’s Annexin V Apoptosis Detection Kit is designed to rapidly analyse different stages of apoptosis and cell death in single cell suspensions.

Annexin V binds specifically to phosphatidylserine (PS) which is located to the inner side of the cytoplasmic membrane in intact cells. But after initiation of apoptosis PS becomes rapidly translocated to the outer side of the cytoplasmic membrane where it is thought to play an important role in macrophage recognition, thus allowing the apoptotic cells to become rapidly phagocytosed.

The binding of annexin V to PS is Ca2+-dependent and therefore, a specific Ca2+-containing reaction buffer is needed during the binding process. At low PS concentrations a binding ratio of eight annexin V molecules to one PS has been reported making annexin V-conjugates ideal for identifying membrane changes associated with early apoptotic events.

In late apoptosis/necrosis cells loose their membrane integrity which allows further annexin V binding to PS located to the inner cell membrane. Therefore, MabTag’s Annexin V Apoptosis Detection Kit contains a viability dye such as propidium iodide (PI). This DNA-intercalating dye can only pass membranes which lost their integrity and therefore, allow the discrimination of early apoptotic cells (Annexin V+/PI-) from late apoptotic/necrotic cells (Annexin V+/PI+).

Cells were treated with UV for 30 minutes. After an incubation of 4 hours they were subsequently stained according to the procedure described below.

 Annexin-V vs. PI

1. viable cells (double-negative)

2. early apoptotic cells (annexin V-positive/PI-negative)

3. late apoptotic/necrotic cells (double-positive)

Characteristics of Fluorochromes

Fluorochrom

Absorption peak (nm)

Emission peak (nm)

Excitation at (nm)

FITC

495

519

488

APC

650

660

633

Propidium iodide

536

617

488

Reagents provided

500 l Annexin V FITC / APC / Biotin conjugate; (with 1% BSA / 0.1% NaN3 as preservative)

500 l propidium iodide [20 g/ml]

10 ml Annexin V binding buffer (10x concentrate)

 

Staining procedure

1. Dilute an appropriate amount of 10x Annexin-V binding buffer with deionized or distilled water in 1:10 ratio

2. Wash cells with culture medium or PBS

3. Resuspend 104 – 106 cells in 90 l of diluted (1x) Annexin-V binding buffer.

4. Add 5 l of Annexin-V conjugate and 5 l of propidium iodide solution.

5. Incubate 20 minutes in the dark.

6. Add 400 l of Annexin V binding buffer (1x).

7. Centrifuge at 400 x g for 5 minutes.

8. Resuspend cells in approx. 500 l Annexin-V binding buffer (1x) and analyze sample by flow cytometry or fluorescence microscopy.