Conjugados de anexina V para la detección de apoptosis
Conjugados de anexina V para la detección de apoptosis
Citas y referencias (30)
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.
Have Questions?
Cambiar vistabuttonViewtableView
Número de catálogoExcitación/emisiónLíneas láser del citómetro de flujoConjugado
A23204650/665633-637Alexa Fluor 647
A13201495/519488Alexa Fluor 488
A13199494/518488FITC
A13202578/603532, 561Alexa Fluor 568
A13203590/617532Alexa Fluor 594
A13204Biotina X
A23202346/442UVAlexa Fluor 350
A35108555/565532, 561Alexa Fluor 555
A35109679/702633-637Alexa Fluor 680
A35110650/660633-637APC (Aloficocianina)
A35111565/578488, 532, 561PE
A35122410/455405Pacific Blue
Número de catálogo A23204
Precio (CLP)
447.525
Each
Añadir al carro de la compra
Excitación/emisión:
650/665
Líneas láser del citómetro de flujo:
633-637
Conjugado:
Alexa Fluor 647
Precio (CLP)
447.525
Each
Añadir al carro de la compra
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
ColorRojo lejano
DescripciónConjugado de anexina V, Alexa Fluor 647
Excitación/emisión650/665
Líneas láser del citómetro de flujo633-637
Para utilizar con (equipo)Citómetro de flujo
Contenido del kitContiene 1 vial de conjugado de anexina V, Alexa Fluor 647.
N.º de reacciones100
Tipo de productoConjugado de anexina V
Cantidad500 μL
Condiciones de envíoHielo húmedo
ConjugadoAlexa Fluor 647
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 (30)

Citations & References
Abstract
The influenza A virus PB1-F2 protein targets the inner mitochondrial membrane via a predicted basic amphipathic helix that disrupts mitochondrial function.
Authors:Gibbs JS, Malide D, Hornung F, Bennink JR, Yewdell JW
Journal:J Virol
PubMed ID:12805420
'The 11th influenza A virus gene product is an 87-amino-acid protein provisionally named PB1-F2 (because it is encoded by an open reading frame overlapping the PB1 open reading frame). A significant fraction of PB1-F2 localizes to the inner mitochondrial membrane in influenza A virus-infected cells. PB1-F2 appears to enhance virus-induced ... More
High-resolution mapping reveals topologically distinct cellular pools of phosphatidylserine.
Authors:Fairn GD, Schieber NL, Ariotti N, Murphy S, Kuerschner L, Webb RI, Grinstein S, Parton RG,
Journal:J Cell Biol
PubMed ID:21788369
'Phosphatidylserine (PS) plays a central role in cell signaling and in the biosynthesis of other lipids. To date, however, the subcellular distribution and transmembrane topology of this crucial phospholipid remain ill-defined. We transfected cells with a GFP-tagged C2 domain of lactadherin to detect by light and electron microscopy PS exposed ... More
Nuclear relocation of the nephrin and CD2AP-binding protein dendrin promotes apoptosis of podocytes.
Authors:Asanuma K, Campbell KN, Kim K, Faul C, Mundel P
Journal:Proc Natl Acad Sci U S A
PubMed ID:17537921
'Kidney podocytes and their slit diaphragms (SDs) form the final barrier to urinary protein loss. There is mounting evidence that SD proteins also participate in intracellular signaling pathways. The SD protein nephrin serves as a component of a signaling complex that directly links podocyte junctional integrity to actin cytoskeletal dynamics. ... More
Transcriptional up-regulation of ULK1 by ATF4 contributes to cancer cell survival.
Authors:Pike LR, Singleton DC, Buffa F, Abramczyk O, Phadwal K, Li JL, Simon AK, Murray JT, Harris AL
Journal:Biochem J
PubMed ID:23078367
'Hypoxia in the microenvironment of many solid tumours is an important determinant of malignant progression. The ISR (integrated stress response) protects cells from the ER (endoplasmic reticulum) stress caused by severe hypoxia. Likewise, autophagy is a mechanism by which cancer cells can evade hypoxic cell death. In the present paper ... More
Phosphorylation of inositol 1,4,5-trisphosphate receptors by protein kinase B/Akt inhibits Ca2+ release and apoptosis.
Authors:Szado T, Vanderheyden V, Parys JB, De Smedt H, Rietdorf K, Kotelevets L, Chastre E, Khan F, Landegren U, Söderberg O, Bootman MD, Roderick HL,
Journal:Proc Natl Acad Sci U S A
PubMed ID:18250332
'Imbalance of signals that control cell survival and death results in pathologies, including cancer and neurodegeneration. Two pathways that are integral to setting the balance between cell survival and cell death are controlled by lipid-activated protein kinase B (PKB)/Akt and Ca(2+). PKB elicits its effects through the phosphorylation and inactivation ... More
View all Conjugados de anexina V para la detección de apoptosis citations