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DUO92202 Sigma-Aldrich

Duolink® PLA Control Kit - PPI

Control Kit to reliably detect Protein-Protein Interactions (PPI) with Duolink® PLA

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Properties

Related Categories Duolink PLA Accessories, Duolink PLA Technology, Molecular Biology, Protein Interaction, Proteomics More...
application(s)   immunofluorescence: suitable
  proximity ligation assay: suitable
suitability   suitable for fluorescence
shipped in   dry ice
storage temp.   −20°C

Description

Components

This product is comprised of the following:
• Duolink® PLA Control Slides (DUO82202) - two 8-well chamber slides with EGF treated, pre-fixed SK-OV3 cells.
• One vial (15μL) of Rabbit anti-ErbB2/HER2 antibody (HPA001383)
• One vial (25μL) of Mouse anti-EGFR antibody (E3138)

See datasheet for more information.

Other Notes

Other Duolink® reagents needed (30RXN size):
Mouse/Rabbit Duolink® PLA Starter Kit: Red Starter Kit (DUO92101) OR Orange Starter Kit (DUO92102)
OR
PLA Probe pairs (PLUS and MINUS): Anti-Mouse PLUS (DUO92001) and Anti-Rabbit MINUS (DUO92005) or Anti-Mouse MINUS (DUO92004) and Anti-Rabbit PLUS (DUO92002); AND Detection reagents of choice: Orange (DUO92007), Red (DUO92008), FarRed (DUO92013), or Green (DUO92014); AND Wash Buffers (DUO82049) AND Mounting Medium with DAPI (DUO82040).

Application

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

Duolink® PLA Control Kit - PPI is designed to allows users to gain confidence with the Duolink® PLA technology. This Kit contains glass slides with fixed, treated cells and a specific pair of primary antibodies that will detect interacting proteins and reliably generate PLA signals when used with the complementary Duolink® PLA probes and detection reagents. Analysis is carried out using standard immunofluorescence assay equipment.

Specificity
This kit will enable the detection of Protein-Protein interactions. The SK-OV-3 cells contain HER2 and EGFR, which show increased interaction upon treatment with EGF. When combined with appropriate Duolink® PLA Reagents, the Control Kit will enable the successful detection of increased HER2 and EGFR protein interactions in the EGF treated cells.

Application note
In the Duolink® PLA Control Kit, the cells and antibodies are provided, and the conditions have been optimized. When combined with the necessary Duolink® PLA reagents, you will have all you need to successfully perform the PLA experiment and obtain positive results.

The Control kit contains Duolink® PLA Control Slides (Cat. No. DUO82202) - two 8 well glass slides with fixed SK-OV3 Human ovarian cancer cells. The cells were treated with EGF, to induce EGFR and HER2 interaction. The Kit also contains one vial of Anti-HER2 antibody produced in rabbit (HPA001383), and one vial of Monoclonal Anti-EGFR antibody produced in mouse (E3138) – the two primary antibodies you need to detect the two interacting protein

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

• Reliable PLA signal detecting a protein interaction
• Gain confidence with the Duolink® PLA Technology
• Add additional controls to your PLA experiments
• Increased sensitivity due to rolling circle amplification for low abundant targets
• No overexpression or genetic manipulation required
• Relative quantification possible
• Works with any fluorescence microscope and appropriate filters

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

Protocols & Articles

Articles

Duolink® PLA Applications

With Duolink® PLA, you can detect, quantify, and visualize protein-protein interactions, post-translational modifications, and low expression protein detection with an easy-to-execute immunodetection...
Keywords: Acylations, Amplification, Biological processes, Buffers, DNA replication, Electrophoresis, Flow cytometry, Gel electrophoresis, Gene expression, Glycosylations, High performance liquid chromatography, Immunohistochemistry, Mass spectrometry, Metabolism, Microscopy, Phosphorylations, Post translational modifications, Sulfations, Transcription, Transduction, Ubiquitinations

Duolink® References

Duolink proximity ligation assay (PLA) has been used in countless studies in a variety of fields. Browse the following tables to view highlights from the many applications of this technology.
Keywords: Apoptosis, Cancer, Cell biology, Coagulation, DNA replication, Degradations, Epigenetics, Flow cytometry, Gene expression, Growth factors, Immunology, Methylations, Neuroscience, Oxidations, PAGE, Peptide synthesis, Phosphorylations, Polymorphisms, Post translational modifications, Transcription

Protocols

Duolink® PLA Brightfield Protocol

This protocol describes the use of Duolink® PLA reagents for the brightfield detection, visualization, and quantification of individual proteins, protein modifications, and protein interactions in ti...
Keywords: Amplification, Cancer, Dehydration reaction, Gene expression, Immunohistochemistry, Peptide synthesis

Duolink® PLA Product Selection Guide

Getting started with Duolink® PLA is simple. The protocols are very similar to that of IF, IHC, and flow cytometry, however Duolink® PLA offers distinct advantages over these and other traditional me...
Keywords: Amplification, Bacterial conjugations, Buffers, Flow cytometry, Fluorescent microscopy, Gene expression, Immunohistochemistry, Microscopy, Peptide synthesis

Duolink® PLA Troubleshooting Guide

The Duolink® PLA technology for protein detection offers a simple and straightforward protocol that has a set of defined steps. This section will delineate important reminders that will ensure your D...
Keywords: Addition reactions, Amplification, Bacterial conjugations, Buffers, Dehydration reaction, Filtration, Flow cytometry, Immunofluorescence, Immunohistochemistry, Peptide synthesis, Purification, Titrations

How Proximity Ligation Assay (PLA) Works

Duolink® Proximity Ligation Assay (PLA) is a powerful tool that allows in situ detection of endogenous proteins, protein modifications, and protein interactions with high specificity and sensitivity....
Keywords: Amplification, Angiogenesis, Apoptosis, Buffers, Cancer, Catalysis, Cell division, Cell proliferation, Flow cytometry, Fluorescent microscopy, Gene expression, Growth factors, Immunofluorescence, Immunohistochemistry, Ligands, Microscopy, Peptide synthesis, Phosphorylations, Transcription

How to Optimize the Duolink® Proximity Ligation Assay

To successfully execute a Duolink® PLA experiment, there are some necessary considerations to properly prepare for, setup, and execute the assay protocol.
Keywords: Bacterial conjugations, Buffers, Detergents, Flow cytometry, Fluorescent microscopy, Gene expression, Growth factors, Immunofluorescence, Immunohistochemistry, In Situ hybridization, Microscopy, Nucleic acid hybridization, Phosphorylations, Post translational modifications, Sample preparations

Related Content

Duolink® PLA - Protein Detection Technology

Inside every cell, there is a breakthrough waiting to be uncovered. The chance to change the origin of a human disease or discover the tell-tale sign of an irregular protein-protein interaction requi...
Keywords: Amplification, Genetic, Immunofluorescence, Individual protein Immunoprecipitation, Peptide synthesis, Post translational modifications, Protein complex immunoprecipitation, Western blot

Duolink® PLA Custom Services

Duolink® Proximity Ligation Assay (PLA) is a powerful tool that allows in situ detection of endogenous proteins, protein interactions, and protein modifications with high specificity and sensitivity....
Keywords: Flow cytometry, Genetic, Peptide synthesis

Duolink® PLA Resource Center

Understanding protein function in the cell can create solutions to improve human health. For that reason we created the Duolink® PLA Resource Center, to provide insightful educational resources so yo...
Keywords: Flow cytometry, Peptide synthesis

Search Primary Antibodies for use with Duolink® PLA

The use of high quality, primary antibodies is critical to success for running a Duolink® Proximity Ligation Assay. We offer over 300 PLA-validated antibodies and antibody pair kits, in addition to t...
Keywords: Immunohistochemistry, PAGE, Peptide synthesis

Peer-Reviewed Papers
15

References

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Add to Cart

P3813 Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions
DUO82049 Duolink® In Situ Wash Buffers, Fluorescence
DUO92001 Duolink® In Situ PLA® Probe Anti-Mouse PLUS, Affinity purified Donkey anti-Mouse IgG (H+L)
DUO92002 Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, Affinity purified Donkey anti-Rabbit IgG (H+L)
DUO92004 Duolink® In Situ PLA® Probe Anti-Mouse MINUS, Affinity purified Donkey anti-Mouse IgG (H+L)
DUO92005 Duolink® In Situ PLA® Probe Anti-Rabbit MINUS, Affinity purified Donkey anti-Rabbit IgG (H+L)
DUO92007 Duolink® In Situ Detection Reagents Orange
DUO92008 Duolink® In Situ Detection Reagents Red
DUO92013 Duolink® In Situ Detection Reagents FarRed
DUO92014 Duolink® In Situ Detection Reagents Green
DUO82040 Duolink® In Situ Mounting Medium with DAPI
DUO92101 Duolink® In Situ Red Starter Kit Mouse/Rabbit
DUO92102 Duolink® In Situ Orange Starter Kit Mouse/Rabbit
T8787 Triton X-100, for molecular biology

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