Annexin V Apoptosis Detection Kit FITC

Catalog No. ABIN2669905

Product Details

Target: Annexin V (ANXA5) (Annexin A5 (ANXA5))

Reactivity: Human, Rodents

Detection Method: Fluorometric

Conjugate: FITC

Application: Flow Cytometry (FACS)

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

Sample Type: Blood, Cell Culture Cells

Characteristics: Staining cells simultaneously with Annexin V-FITC and the non-vital propidium ioide allows (bivariate analysis) the discrimination of intact cells (Annexin V-FITC negative, PI negative), early apoptotic (Annexin V-FITC positive, PI negative) and late apoptotic or necrotic cells (Annexin V-FITC positive, PI positive).

Components:

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

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

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.

Sample Volume: 5 μL

Assay Time: < 1 h

Protocol: Staining cells protocol with Annexin-FITC. 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-FITC and 5 μL of PI, 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.

Reagent Preparation:

• AnnexinV FITC is ready to use.
• PI 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

Handling

Preservative: Sodium azide

Precaution of Use: 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

Handling Advice: Light exposure should be avoided. Use dim light during handling, incubation with cells and prior to analysis.

Storage: 4 °C

References

Cottone, Capobianco, Gualteroni, Perrotta, Bianchi, Rovere-Querini, Manfredi: "5-Fluorouracil causes leukocytes attraction in the peritoneal cavity by activating autophagy and HMGB1 release in colon carcinoma cells." in: International journal of cancer. Journal international du cancer, Vol. 136, Issue 6, pp. 1381-9, (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." in: 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." in: 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).

Ocio, Fernández-Lázaro, San-Segundo, López-Corral, Corchete, Gutiérrez, Garayoa, Paíno, García-Gómez, Delgado, Montero, Díaz-Rodríguez, Mateos, Pandiella, Couto, Wang, Bjorklund, San-Miguel: "In vivo murine model of acquired resistance in myeloma reveals differential mechanisms for lenalidomide and pomalidomide in combination with dexamethasone." in: Leukemia, Vol. 29, Issue 3, pp. 705-14, (2015) (PubMed).

Acebes-Huerta, Huergo-Zapico, Gonzalez-Rodriguez, Fernandez-Guizan, Payer, López-Soto, Gonzalez: "Lenalidomide induces immunomodulation in chronic lymphocytic leukemia and enhances antitumor immune responses mediated by NK and CD4 T cells." in: BioMed research international, Vol. 2014, pp. 265840, (2014) (PubMed).

Germano, Rapa, Volante, Lo Buono, Carturan, Berruti, Terzolo, Papotti: "Cytotoxic activity of gemcitabine, alone or in combination with mitotane, in adrenocortical carcinoma cell lines." in: Molecular and cellular endocrinology, Vol. 382, Issue 1, pp. 1-7, (2013) (PubMed).

Target Details

Target: Annexin V (ANXA5) (Annexin A5 (ANXA5))

Alternative Name: Annexin V (ANXA5 Products)

Synonyms: anx Kit, anx5 Kit, ANX V Kit, anxa5 Kit, cb989 Kit, wu:fa98f06 Kit, wu:fj10f10 Kit, MGC89158 Kit, ANX5 Kit, ENX2 Kit, PP4 Kit, RPRGL3 Kit, Anx5 Kit, R74653 Kit, LC5 Kit, enx2 Kit, annexin A5 Kit, annexin A5b Kit, Annexin A5 Kit, annexin A5 L homeolog Kit, ANXA5 Kit, anxa5b Kit, anxa5 Kit, Anxa5 Kit, anxa5.L Kit

Molecular Weight: 35 kDa