Annexin V Apoptosis Detection Kit PE

N° du produit ABIN2669906

Aperçu rapide pour Annexin V Apoptosis Detection Kit PE (ABIN2669906)

Antigène: Annexin V (ANXA5) (Annexin A5 (ANXA5))

Reactivité: Chemical

Méthode de détection: Fluorometric

Application: Flow Cytometry (FACS)

Type d'échantillon: Blood, Cell Culture Cells

Détail du produit

Conjugué: PE

Fonction: The kit can identify and quantitate apoptotic cells on a single-cell basis by flow cytometry.

Attributs du produit: Staining cells simultaneously with Annexin V-PE and the non-vital 7-Amino-Actinomycin D (far red fluorescence) allows (bivariate analysis) the discrimination of intact cells (Annexin V-PE negative, 7-AAD negative), early apoptotic (Annexin V-PE positive, 7-AAD negative) and late apoptotic or necrotic cells (Annexin V-PE positive, 7-AAD positive).

Ingrédients:

• 100 Tests of AnnexinV FITC
• 100 Tests of PI
• 10X Binding Buffer

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Commentaires:

Samples were tested on Flow Cytometry Induce apoptosis in cells using the desired method is not included in this time. For instance, Jurkat cells (T-cell leukemia, human) treated with 6 μM camptothecin for four hours.

Volume d'échantillon: 5 μL

Durée du test: < 1 h

Protocole: Staining cells protocol with Annexin-PE. Flow Cytometry
1. Prepare Annexin V Binding Buffer: 10 mM Hepes/NaOH ( pH 7,4) 140 mM NaCl, 2,5 mM CaCl2. .
2. Induce apoptosis in cells using the desired method. A negative control should be prepared by untreated cells, that is used to define the basal level of apoptotic and necrotic or dead cells.
3. Harvest the cells after the apoptosis induction and wash in temperate phosphate-buffered saline (PBS).
4. Wash cells twice with temperate PBS and resuspend cells in 1 X Annexin-binding buffer at a concentration 1 x 106 cells/mL.
5. Add 5 μL of the Annexin V-PE and 5 μL of 7-AAD, to each 100 μL of cell suspension.
6. Incubate the cells at room temperature for 15 minutes at room temperature (25 °C) in the dark.
7. After incubation period, add 400 μL of 1X Annexin-binding buffer. Analyze by flow cytometry within one hour.

Préparation des réactifs:

• AnnexinV PE is ready to use.
• 7AAD is ready to use.
• Prepare Annexin V Binding Buffer
10 mM Hepes/NaOH (pH 7,4) 140 mM NaCl, 2,5 mM CaCl2.

Restrictions: For Research Use only

Stockage

Agent conservateur: Sodium azide

Précaution d'utilisation: Reagents contain sodium azide. Sodium azide under acid conditions yields hydrazoic acid, an extremely toxic compound. Azide compounds should be diluted with running water before being discarded. These conditions are recommended to avoid deposits in plumbing where explosive conditions may develop.
Do not pipet by mouth.
Samples should be handled as if capable of transmitting infection. Appropriate disposal methods should be used.
The sample preparation procedure employs a fixative (formaldehyde). Contact is to be avoided with skin or mucous membranes

Conseil sur la manipulation: Light exposure should be avoided. Use dim light during handling, incubation with cells and prior to analysis.

Stock: 4 °C

Référence

Mazzeo, Calvo, Alonso, Mérida, Izquierdo: "Protein kinase D1/2 is involved in the maturation of multivesicular bodies and secretion of exosomes in T and B lymphocytes." dans: Cell death and differentiation, (2015) (PubMed).

De Miguel, Gallego-Lleyda, Galan-Malo, Rodriguez-Vigil, Marzo, Anel, Martinez-Lostao: "Immunotherapy with liposome-bound TRAIL overcomes partial protection to soluble TRAIL-induced apoptosis offered by down-regulation of Bim in leukemic cells." dans: Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico, Vol. 17, Issue 8, pp. 657-67, (2015) (PubMed).

Ramírez-Labrada, López-Royuela, Jarauta, Galán-Malo, Azaceta, Palomera, Pardo, Anel, Marzo, Naval: "Two death pathways induced by sorafenib in myeloma cells: Puma-mediated apoptosis and necroptosis." dans: Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico, Vol. 17, Issue 2, pp. 121-32, (2015) (PubMed).

Détail du antigène

Antigène: Annexin V (ANXA5) (Annexin A5 (ANXA5))

Autre désignation: Annexin V

Poids moléculaire: 35 kDa