Merck

DUO92012

Sigma-Aldrich

Duolink® In Situ Detection Reagents Brightfield

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Sinónimos:
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent

Línea del producto

Duolink®

envase

pkg of Box A (-20°C)
pkg of Box B (2-8°C)

técnicas

proximity ligation assay: suitable

idoneidad

suitable for brightfield

temp. de almacenamiento

−20°C

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Este artículo
DUO92007DUO82049DUO92010
suitability

suitable for brightfield

suitability

suitable for fluorescence

suitability

suitable for fluorescence

suitability

suitable for brightfield, suitable for fluorescence

storage temp.

−20°C

storage temp.

−20°C

storage temp.

20-25°C

storage temp.

−20°C

product line

Duolink®

product line

Duolink®

product line

Duolink®

product line

Duolink®

packaging

pkg of Box A (-20°C)

packaging

-

packaging

-

packaging

-

Aplicación

Duolink®proximity ligation assay(PLA®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

Follow the Duolink® In Situ Brightfield Protocol to use this product.

Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink® PLA probes. Analysis is carried out using HRP for brightfield detection.
Specificity
Brightfield detection reagents are used for samples with significant autofluorescence.

Application Note
Two primary antibodies raised in different species are needed. Test your primary antibodies (IgG-class, mono- or polyclonal) in a standard immunofluorescence (IF), immunohistochemistry (IHC) or immunocytochemistry (ICC) assay to determine the optimal fixation, blocking, and titer conditions. Duolink® in situ reagents are suitable for use on fixed cells, cytospin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

View full Duolink® product list

Características y beneficios

  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Single molecule sensitivity due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Quicker and simpler than FRET
  • Increased accuracy compared to co-IP
  • Publication-ready results

Componentes

This product is comprised of the following:
Box A:
  • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
  • 1x Ligase (1 unit/μL)
  • 5x Amplification - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase
  • 5x Detection Brightfield - Contains oligonucleotide probes labeled with horseradish peroxidase (HRP) that hybridize to the RCA product
  • 1x Polymerase (10 units/μL)

Box B:
  • 1x Hydrogen Peroxide - Contains 0.3% hydrogen peroxide
  • Substrate Reagents A-D - Contains all substrate components needed for HRP enzymatic reaction
  • 1x Nuclear Stain - Contains Mayer′s hematoxylin solution for cell nuclei staining
See datasheet for more information.

Not included in Detection kit:

Primary antibodies, PLA probes, wash buffers, mounting medium

Otras notas

Storage/Stability
Store Box A and all of its components at -20°C. The enzymes should be kept cold (-20°C) at all times; Use a freezing block when removing them from the freezer.
Store Box B and all of its components at 2-8°C.

Información legal

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

Palabra de señalización

Danger

Clasificaciones de peligro

Acute Tox. 3 Dermal - Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 3 - Eye Dam. 1 - Flam. Liq. 2 - Muta. 2 - Skin Irrit. 2 - Skin Sens. 1

Código de clase de almacenamiento

3 - Flammable liquids

Clase de riesgo para el agua (WGK)

WGK 3

Punto de inflamabilidad (°F)

42.8 °F

Punto de inflamabilidad (°C)

6 °C


Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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Ejemplo

T1503
Referencia del producto
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25G
Tamaño del envase/cantidad

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Artículos

Learn how Proximity Ligation Assay technology works and how the protein-protein interaction control kit can confirm in situ detection of EGF-induced EGFR-HER2 dimerization.

Things to consider for preparation, setup and execution of the Duolink® assay protocol

Support information including tips and tricks, frequently asked questions, and basic troubleshooting.

Protocolos

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Contenido relacionado

Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay

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