MilliporeSigma
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DUO92008

Sigma-Aldrich

Duolink® In Situ Detection Reagents Red

Synonym(s):
Anti-HOKPP1, Anti-HOKPP
NACRES:
NA.32

Quality Level

product line

Duolink®

technique(s)

proximity ligation assay: suitable

fluorescence

λex 594 nm; λem 624 nm (Texas Red®, Zeiss Filter set 31)

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

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.

Follow the Duolink® In Situ Fluorescence Protocol to use 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
Red fluorescence detection reagents are often used with Texas Red® filter.

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

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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

Components

This product is comprised of the following:
  • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
  • 1x Ligase (1 unit/μL)
  • 1x Polymerase (10 units/μL)
5x Amplification Red - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase. It also contains oligonucleotide probes labeled with a fluorophore that hybridize to the RCA product.
See datasheet for more information.

Not included in Detection kit:

Primary antibodies, PLA probes, wash buffers, mounting medium

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC
Texas Red is a registered trademark of Life Technologies

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Articles

Detection of Protein-protein Interactions by in situ PLA

Proteins are complex biological molecules essential for cellular structure and functions. The majority of proteins commonly interact with various molecules, including other proteins, in order to exert their functions.

Duolink® PLA Fluorescence Protocol

Protocol for immunofluorescent detection of proteins in cells and tissue

How to Optimize the Duolink® Proximity Ligation Assay

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

Duolink® References

Find Duolink references based on the type of method used, post translational modification detected, and research focus.

See All

Protocols

Duolink® In Situ Short Instructions - Fluorescence

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

Related Content

Duolink® PLA Applications

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

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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