Annexin V Apoptosis Detection Kit FITC

Details for Product No. ABIN2669905, Supplier: Log in to see
Antigen
  • anx
  • anx5
  • ANX V
  • anxa5
  • cb989
  • wu:fa98f06
  • wu:fj10f10
  • MGC89158
  • ANX5
  • ENX2
  • PP4
  • RPRGL3
  • Anx5
  • R74653
  • LC5
  • enx2
  • annexin A5
  • annexin A5b
  • Annexin A5
  • annexin 5
  • annexin A5-like
  • ANXA5
  • anxa5b
  • anxa5
  • LOC100220003
  • LOC100305020
  • LOC100342904
  • Anxa5
  • LOC100521982
Alternatives
Human Annexin V ELISA Kit
Reactivity
Human, Rodentia
16
16
15
14
13
8
7
7
4
2
2
2
2
1
1
1
Conjugate
FITC
Application
Flow Cytometry (FACS)
Options
Supplier
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Supplier Product No.
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Purpose The kit can identify and quantitate apoptotic cells on a single-cell basis by flow cytometry.
Sample Type Blood, Cell Culture Cells
Detection Method Fluorometric
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
Material not included
  • pipettes
  • ,
  • tubes
  • ,
  • flow cytometry machine
Alternative Name Annexin V (ANXA5 ELISA Kit Abstract)
Molecular Weight 35 kDa
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
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
Product cited in: 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).

Background publications Herrero-Martín, Osuna, Ordóñez, Sevillano, Martins, Mackintosh, Campos, Madoz-Gúrpide, Otero-Motta, Caballero, Amaral, Wai, Braun, Eisenacher, Schaefer, Poremba, de Alava: "Stable interference of EWS-FLI1 in an Ewing sarcoma cell line impairs IGF-1/IGF-1R signalling and reveals TOPK as a new target." in: British journal of cancer, Vol. 101, Issue 1, pp. 80-90, 2009 (PubMed).

Pérez-Andrés, Benito, Rodríguez-Fernández, Corradetti, Primo, Manzano, Orfao, Criado: "Bisursodeoxycholate(ethylenediamine)platinum(II): a new autofluorescent compound. Cytotoxic activity and cell cycle analysis in ovarian and hematological cell lines." in: Dalton transactions (Cambridge, England : 2003), Issue 44, pp. 6159-64, 2008 (PubMed).

Darzynkiewicz, Bedner, Traganos: "Difficulties and pitfalls in analysis of apoptosis." in: Methods in cell biology, Vol. 63, pp. 527-46, 2001 (PubMed).

Homburg, de Haas, von dem Borne, Verhoeven, Reutelingsperger, Roos: "Human neutrophils lose their surface Fc gamma RIII and acquire Annexin V binding sites during apoptosis in vitro." in: Blood, Vol. 85, Issue 2, pp. 532-40, 1995 (PubMed).

Vermes, Haanen, Steffens-Nakken, Reutelingsperger: "A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V." in: Journal of immunological methods, Vol. 184, Issue 1, pp. 39-51, 1995 (PubMed).

Koopman, Reutelingsperger, Kuijten, Keehnen, Pals, van Oers: "Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis." in: Blood, Vol. 84, Issue 5, pp. 1415-20, 1994 (PubMed).

Fadok, Voelker, Campbell, Cohen, Bratton, Henson: "Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 148, Issue 7, pp. 2207-16, 1992 (PubMed).