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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

Related Categories

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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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 Code

10 - Combustible liquids

WGK

WGK 3

Certificates of Analysis (COA)

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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)
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)
Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction
Mendez R, et al.
Bio-protocol, 7(21) (2017)
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
M Aubele et al.
British journal of cancer, 103(5), 663-667 (2010-08-12)
Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression
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