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DUO82049

Sigma-Aldrich

Duolink® In Situ Wash Buffers, Fluorescence

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Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
NACRES:
NA.32

material

packing (powdered buffer pouches)

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for fluorescence

storage temp.

20-25°C

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DUO82047DUO92010DUO92009
product line

Duolink®

product line

Duolink®

product line

Duolink®

product line

Duolink®

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

suitability

suitable for fluorescence

suitability

suitable for brightfield

suitability

suitable for brightfield, suitable for fluorescence

suitability

suitable for brightfield, suitable for fluorescence

storage temp.

20-25°C

storage temp.

20-25°C

storage temp.

−20°C

storage temp.

−20°C

material

packing (powdered buffer pouches)

material

packing (powdered buffer pouches)

material

-

material

-

Application

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.

Use the Duolink® In Situ Fluorescence Protocol for this product. A set of short instructionsis also available.

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 standard immunofluorescence assay equipment.
Specificity
Duolink® In Situ fluorescence applications use two wash buffers. Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents. See datasheet for more information.

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®PLA 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.

Linkage
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View full Duolink® product list
Duolink® PLA 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).

Features and Benefits

  • 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

Legal Information

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

Storage Class

10 - Combustible liquids

wgk_germany

WGK 3


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Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction
Mendez R, et al.
Bio-protocol, 7(21) (2017)
Detection of Receptor Heteromerization Using In Situ Proximity Ligation Assay
Gomes I, et al.
Current Protocols in Pharmacology / Editorial Board, S.J. Enna (editor-in-chief) ... [Et al.], 2-16 (2016)
Increasing the Receptor Tyrosine Kinase EphB2 Prevents Amyloid-?-induced Depletion of Cell Surface Glutamate Receptors by a Mechanism That Requires the PDZ-binding Motif of EphB2 and Neuronal Activity
Miyamoto T, et al.
The Journal of Biological Chemistry (2015)
Xiaofan Li et al.
PLoS pathogens, 13(3), e1006249-e1006249 (2017-03-02)
Trials to reintroduce chloroquine into regions of Africa where P. falciparum has regained susceptibility to chloroquine are underway. However, there are long-standing concerns about whether chloroquine increases lytic-replication of Epstein-Barr virus (EBV), thereby contributing to the development of endemic Burkitt
Xiaofan Li et al.
Journal of virology, 93(17) (2019-06-14)
Herpesviruses are ubiquitous, and infection by some, like Epstein-Barr virus (EBV), is nearly universal. To persist, EBV must periodically switch from a latent to a replicative/lytic phase. This productive phase is responsible for most herpesvirus-associated diseases. EBV encodes a latency-to-lytic

Articles

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.

Protocols

Duolink® PLA Multicolor Detection Protocol

This page details the Duolink® In Situ Short Protocol for fluorescence detection

This protocol describes how to perform immunofluorescent detection of proteins in cells and tissue.

Related Content

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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