Reactivo ReadyProbes™ para potenciador de señal Image-iT™ FX

Citas y referencias (10)

Invitrogen™

Reactivo ReadyProbes™ para potenciador de señal Image-iT™ FX

El reactivo ReadyProbes™ para potenciador de señal Image-iT™ reduce drásticamente la fluorescencia de fondo, lo que resulta en una tinciónMás información

Número de catálogo	Cantidad
R37107	6 x 2.5 mL kit

Número de catálogo R37107

Precio (CLP)

363.109

Añadir al carro de la compra

Cantidad:

6 x 2.5 mL kit

Precio (CLP)

363.109

Añadir al carro de la compra

El reactivo ReadyProbes™ para potenciador de señal Image-iT™ reduce drásticamente la fluorescencia de fondo, lo que resulta en una tinción no específica durante la detección secundaria con conjugados de estreptavidina o anticuerpos secundarios. Este reactivo se aplica directamente a los portaobjetos o cubreobjetos que contienen muestras de tejido o células fijas y permeabilizadas antes de la tinción con sondas fluorescentes.

• Protocolo sencillo: aplique el potenciador a partir de una solución preparada en un frasco cuentagotas
• Compatible con los otros pasos de su protocolo de inmunotinción
• Eficaz con muchas etiquetas fluorescentes
• Estables a temperatura ambiente: manténgalo a mano en su zona de trabajo o área de cultivo celular

Consulte otros reactivos ReadyProbes™ para la tinción celular
Descubra más herramientas para la obtención de imágenes celulares fijas

Aplicaciones de imágenes celulares
El reactivo potenciador de señal Image-iT™ FX es un producto altamente eficaz para bloquear la tinción de fondo que resulta de las interacciones inespecíficas de una amplia variedad de colorantes fluorescentes con componentes celulares y tisulares. La tinción de fondo observada con conjugados fluorescentes de estreptavidina y conjugados de anticuerpos se elimina en gran medida cuando el reactivo potenciador de señal Image-iT™ FX se aplica a células fijas y permeables antes de la tinción.

Sugerencias de uso
• Aplique el reactivo potenciador de señal Image-iT™ FX directamente en los portaobjetos o cubreobjetos que contengan muestras de células o tejidos antes de la inmunotinción.
• Después del paso de fijación/permeabilización, simplemente lave y añada suficientes gotas para cubrir la muestra; luego incube durante 30 minutos y continúe con el protocolo de tinción normal.
• Se pueden realizar pasos de bloqueo adicionales después del bloqueo con el reactivo potenciador de señal Image-iT™ ReadyProbes™, según sea necesario

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

FormatoBottle(s)

Línea de productosImage-iT, ReadyProbes

Cantidad6 x 2.5 mL kit

Tipo de productoReactivo potenciador de señal FX

Tipo de soluciónPotenciadores de señal

Unit SizeEach

Contenido y almacenamiento

6 × Frascos cuentagotas de 2,5 ml

Almacenar a ≤25 °C.

Preguntas frecuentes

I used Image-iT FX Signal Enhancer solution to get rid of nonspecific nuclear labeling with Alexa Fluor 568 secondary antibody, but I also saw a significant reduction in my specific mitochondrial antibody labeling. Why is this and what can I do?

The Image-iT FX Signal Enhancer reduces non-specific binding of dye conjugates by blocking positively-charged areas of cells or tissues that attract negatively-charged dyes. In cells after fixation, some positively-charged structures are the nuclei and mitochondria. Thus, you would expect to see a reduction in both mitochondrial and nuclear signal. The lower signal you see afterward is the specific mitochondrial signal; the fluorescence that was lost was the non-specific labeling.

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

Can I use the Image-iT FX Signal Enhancer instead of my normal blocking solution (BSA or serum)?

No. Image-iT FX Signal Enhancer is not a protein blocker, like BSA, normal serum, or other commercial antibody blockers. Use it as a separate step to block non-specific charge-based binding of dyes to cellular components.

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

I am labeling fixed and permeabilized cultured cells with an Alexa Fluor secondary antibody. My secondary-only control is showing nuclear and mitochondrial labeling, even though I did a thorough protein blocking and tried a concentration range. What can be done to minimize this non-specific binding?

This is most likely charge-based binding due to interactions of the charge on the dyes with cellular components of opposite charge. This can be blocked by using the Image-iT FX Signal Enhancer Solution which eliminates non-specific binding due to charge. The Signal Enhancer is applied as a pre-blocking step and your regular blocking regimen should be used to limit non-specific binding due to protein-protein interactions. Signal Enhancer cannot be used on live cells.

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

Can I use the ReadyProbes reagents for flow cytometry?

This is not recommended. The ReadyProbes reagents were developed for imaging applications whereas the Ready Flow reagents were optimized for flow cytometry.

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

I am observing high background when I analyze my click-labeled samples. What is causing this and what can I do to reduce the background?

The click reaction is very selective between an azide and alkyne. No other side reactions are possible in a biological system. Any non-specific background is due to non-covalent binding of the dye to various cellular components. The Select FX Signal Enhancer is not effective at reducing non-specific charge-based binding of dyes following the click reaction; we do not recommend its use with the Click-iT detection reagents. The best method to reduce background is to increase the number of BSA washes. You should always do a no-dye or no-click reaction control under the same processing and detection conditions to verify that the background is actually due to the dye and not autofluorescence. You can also perform the complete click reaction on a carrier solvent-only, no EdU or no-EU control to verify the specificity of the click reaction signal.

Find additional tips, troubleshooting help, and resources within our Cell Viability, Proliferation, Cryopreservation, and Apoptosis Support Center.

View all Reactivo ReadyProbes™ para potenciador de señal Image-iT™ FX FAQs

Citations & References (10)

Citations & References

Abstract

Functional scaffold-free 3-D cardiac microtissues: a novel model for the investigation of heart cells.

Authors:Desroches BR, Zhang P, Choi BR, King ME, Maldonado AE, Li W, Rago A, Liu G, Nath N, Hartmann KM, Yang B, Koren G, Morgan JR, Mende U,

Journal:Am J Physiol Heart Circ Physiol

PubMed ID:22427522

To bridge the gap between two-dimensional cell culture and tissue, various three-dimensional (3-D) cell culture approaches have been developed for the investigation of cardiac myocytes (CMs) and cardiac fibroblasts (CFs). However, several limitations still exist. This study was designed to develop a cardiac 3-D culture model with a scaffold-free technology ... More

Generation of an induced pluripotent stem cell line (TRNDi008-A) from a Hunter syndrome patient carrying a hemizygous 208insC mutation in the IDS gene.

Authors:Hong J, Xu M, Li R, Cheng YS, Kouznetsova J, Beers J, Liu C, Zou J, Zheng W

Journal:Stem Cell Res

PubMed ID:31071499

'Mucopolysaccharidosis Type II (MPS II), also known as Hunter syndrome, is a rare X-linked genetic disease caused by mutations in the IDS gene encoding iduronate 2-sulfatase (I2S). This is a multisystem disorder with significant variation in symptoms. Here, we document a human induced pluripotent stem cell (iPSC) line generated from ... More

Three-dimensional residual channel attention networks denoise and sharpen fluorescence microscopy image volumes.

Authors:

Journal:Nat Methods

PubMed ID:34059829

The aberrant upregulation of exon 10-inclusive SREK1 through SRSF10 acts as an oncogenic driver in human hepatocellular carcinoma.

Authors:

Journal:Nat Commun

PubMed ID:35296659

HIV-infected macrophages resist efficient NK cell-mediated killing while preserving inflammatory cytokine responses.

Authors:

Journal:Cell Host Microbe

PubMed ID:33571449

View all Reactivo ReadyProbes™ para potenciador de señal Image-iT™ FX citations