Conjugados de anexina V para la detección de apoptosis
Conjugados de anexina V para la detección de apoptosis
Citas y referencias (9)
Invitrogen™

Conjugados de anexina V para la detección de apoptosis

Detecte las primeras etapas de la apoptosis con la anexina V independiente Alexa Fluor, APC, Pacific Blue, PE, FITC y conjugados de biotina mediante citometría de flujo.
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Número de catálogoExcitación/emisiónLíneas láser del citómetro de flujoConjugado
A35122410/455405Pacific Blue
A13201495/519488Alexa Fluor 488
A13199494/518488FITC
A13202578/603532, 561Alexa Fluor 568
A23202346/442UVAlexa Fluor 350
A23204650/665633-637Alexa Fluor 647
A35108555/565532, 561Alexa Fluor 555
A35109679/702633-637Alexa Fluor 680
A35110650/660633-637APC (Aloficocianina)
A35111565/578488, 532, 561PE
Número de catálogo A35122
Precio (USD)
-
Excitación/emisión:
410/455
Líneas láser del citómetro de flujo:
405
Conjugado:
Pacific Blue
Consiga una detección rápida y fiable de la apoptosis celular temprana con conjugados independientes de anexina V para la detección de apoptosis. Los conjugados de anexina V ofrecen una diferencia de hasta 100 veces en la intensidad de la señal de fluorescencia entre células apoptóticas y no apoptóticas mediante citometría de flujo.
La anexina V tiene una alta afinidad a la fosfatidilserina (PS), que se expone en el recubrimiento externo de las células que experimentan apoptosis. Debido a esta afinidad, los reactivos de anexina V etiquetados fluorescentemente se utilizan comúnmente en la investigación de la apoptosis.

Los conjugados de anexina V proporcionan métodos de detección rápidos y fiables para estudiar la externalización de la fosfatidilserina, un indicador de las etapas intermedias de la apoptosis. La diferencia en la intensidad de la fluorescencia entre las células apoptóticas y no apoptóticas teñidas con nuestros conjugados de anexina V fluorescente, medida por citometría de flujo, suele ser de aproximadamente 100 veces.

En colaboración con Nexins Research BV, proporcionamos los mejores y más brillantes conjugados de anexina V disponibles, incluidos los conjugados de anexina V Alexa Fluor 350, 488, 555, 568, 594, 647 y 680, así como los conjugados de anexina V APC, biotina-X, FITC, Pacific Blue y PE. Los conjugados de anexina V altamente fluorescente proporcionan métodos de detección rápidos y fiables para el estudio de la externalización de la fosfatidilserina, uno de los primeros indicadores de apoptosis.

El conjugado de anexina V Pacific Blue es excitable con violeta, lo que lo hace ideal para instrumentos con un láser violeta y para experimentos multicolor que incluyan colorantes verdes o rojos fluorescentes.

Los beneficios de nuestros conjugados de anexina V incluyen:
• Conjugado con los colorantes Invitrogen Alexa Fluor y eFluor para señales más brillantes
• Conjugado para todos los láseres disponibles
• Disponible como reactivos independientes o kits fáciles de usar

La tinción de anexina V para detectar células apoptóticas solo se puede realizar en células y tejidos vivos. Si las muestras se van a fijar después de la tinción, se requieren condiciones específicas para lograr la retención transitoria de la señal. Estas incluyen el uso de un método de fijación a base de aldehídos sin alcohol, el uso de tampones que contengan Ca2+ y evitar los tensioactivos/detergentes. Para su comodidad, también ofrecemos un tampón concentrado de unión a anexina que facilita la unión de la anexina V a la fosfatidilserina en ensayos de apoptosis.

For Research Use Only. Not for use in diagnostic procedures.
Especificaciones
ColorAzul
DescripciónConjugado de anexina V, Pacific Blue para citometría de flujo
Excitación/emisión410/455
Líneas láser del citómetro de flujo405
Para utilizar con (equipo)Citómetro de flujo
Contenido del kitContiene 1 vial de conjugado anexina V, Pacific Blue.
N.º de reacciones100
Tipo de productoConjugado de anexina V
Cantidad500 μL
Condiciones de envíoHielo húmedo
ConjugadoPacific Blue
Unit SizeEach
Contenido y almacenamiento
Almacenar en el refrigerador (2 °C a 8 °C) y proteger de la luz.

Preguntas frecuentes

I want to study apoptosis using an Annexin V conjugate, but with adherent cells via microscopy instead of flow cytometry. Can this be done?

It has been done, but we don‘t recommend it. Both healthy cells and apoptotic cells possess phosphatidylserine on the cell surface, which can be detected with Annexin V, but apoptotic cells have significantly more of it. You can easily tell the difference between these two populations with flow cytometry, because flow cytometers are more sensitive and have a higher throughput. But with a microscope, you cannot always tell the difference, especially for adherent cells. Instead, for microscopy, we recommend a different technique, such as detecting caspases with CellEvent Caspase Detection Reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I trypsinized my adherent cells and labeled with annexin V, and now my flow data is showing a high percentage of apoptotic cells even for control, untreated cells. What is the problem?

Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. Allow the cells to recover for about 30 minutes in optimal cell culture conditions and medium after trypsinizing/scraping so that they can recover their membrane integrity before staining. For lightly adherent cell lines, such as HeLa and NIH 3T3, another option is to use non-enzyme treatments like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I detect annexin V staining in an imaging assay?

Annexin V staining is not typically used in imaging experiments; it is a better reagent for flow cytometry analysis. All cells will stain to some extent, so it can be difficult to distinguish a relatively bright annexin V-stained cell from a dimmer non-apoptotic cell. Caspase activation, detected using our CellEvent Caspase 3/7 or Image-iT LIVE Caspase detection kits, is a better method for detecting apoptosis in an imaging assay.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When should I stain adherent cells with annexin V for flow cytometric analysis? Before or after I trypsinize them?

Trypsinize first and then allow the cells to recover about 30 minutes in optimal cell culture conditions and medium before staining with annexin V conjugates. Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing for annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. For lightly adherent cell lines such as HeLa and NIH 3T3, you could use a less harsh (non-enzymatic) dissociation product like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I fix my cells after annexin V staining?

Yes, this is possible. We have established protocols for annexin V staining combined with intracellular staining of lymphocytes that can be found here. The most important step is to leave some binding buffer in the suspension when fixation is started. Compared to staining of live cells, the intensity of the annexin V signal may be somewhat reduced.

View all Conjugados de anexina V para la detección de apoptosis FAQs

Citations & References (9)

Citations & References
Abstract
Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium, required for virus replication and cell death induction.
Authors:Geng X, Huang C, Qin Y, McCombs JE, Yuan Q, Harry BL, Palmer AE, Xia NS, Xue D,
Journal:Proc Natl Acad Sci U S A
PubMed ID:23091012
'Infection with the hepatitis B virus (HBV) promotes the development of hepatitis, cirrhosis, and hepatocellular carcinoma (HCC) and is a leading cause of morbidity and mortality worldwide. HBV X protein (HBx) is an important effector for HBV pathogenesis, but its cellular targets and acting mechanisms remain elusive. We show here ... More
Live attenuated lentivirus infection elicits polyfunctional simian immunodeficiency virus Gag-specific CD8+ T cells with reduced apoptotic susceptibility in rhesus macaques that control virus replication after challenge with pathogenic SIVmac239.
Authors:Genescà M, Rourke T, Li J, Bost K, Chohan B, McChesney MB, Miller CJ,
Journal:J Immunol
PubMed ID:17878372
'HIV-specific CD8+ T cells that secrete multiple cytokines in response to Ag stimulation are associated with the control of virus replication during chronic HIV infection. To determine whether the presence of polyfunctional CD8+ T cell responses distinguishes protected and unprotected monkeys in a live attenuated lentivirus model, SIV Gag peptide-specific ... More
A novel method for long term bone marrow culture and genetic modification of murine neutrophils via retroviral transduction.
Authors:Zemans RL, Briones N, Young SK, Malcolm KC, Refaeli Y, Downey GP, Worthen GS,
Journal:J Immunol Methods
PubMed ID:19010330
'Neutrophils are a critical component of the innate immune response to invading microbial pathogens. However, an excessive and/or prolonged neutrophil response can result in tissue injury that is thought to underlie the pathogenesis of various inflammatory diseases. The development of novel therapeutic strategies for inflammatory diseases depends on an improved ... More
Modified vaccinia virus Ankara-based vaccine vectors induce apoptosis in dendritic cells draining from the skin via both the extrinsic and intrinsic caspase pathways, preventing efficient antigen presentation.
Authors:Guzman E, Cubillos-Zapata C, Cottingham MG, Gilbert SC, Prentice H, Charleston B, Hope JC,
Journal:J Virol
PubMed ID:22419811
'Dendritic cells (DC) are potent antigen-presenting cells and central to the induction of immune responses following infection or vaccination. The collection of DC migrating from peripheral tissues by cannulation of the afferent lymphatic vessels provides DC which can be used directly ex vivo without extensive in vitro manipulations. We have ... More
The phosphatidylinositol-3-kinase inhibitor NVP-BKM120 overcomes resistance signals derived from microenvironment by regulating the Akt/FoxO3a/Bim axis in chronic lymphocytic leukemia cells.
Authors:Rosich L, Saborit-Villarroya I, López-Guerra M, Xargay-Torrent S, Montraveta A, Aymerich M, Villamor N, Campo E, Pérez-Galán P, Roué G, Colomer D,
Journal:
PubMed ID:23850807
Phosphatidylinositol-3-kinase pathway is constitutively activated in chronic lymphocytic leukemia mainly due to microenvironment signals, including stromal cell interaction and CXCR4 and B-cell receptor activation. Because of the importance of phosphatidylinositol-3-kinase signaling in chronic lymphocytic leukemia, we investigated the activity of the NVP-BKM120, an orally available pan class I phosphatidylinositol-3-kinase inhibitor. ... More
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