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DUO92202

Sigma-Aldrich

Duolink® PLA Control Kit - PPI

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

Synonym(s):
Anti-MSF, Anti-LETS

product line

Duolink®

technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

General description

Duolink® PLA Control Kits are intended for use with Duolink® PLA reagents to ensure proper function of reagents and Duolink® PLA staining technique.

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

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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

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).

Legal Information

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

Storage Class Code

11 - Combustible Solids

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).

Product Information Sheet

Quotes and Ordering

Articles

How Proximity Ligation Assays (PLA) Work

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.

Duolink® References

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

Duolink® PLA Troubleshooting Guide

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

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 tissue and cell samples.

Related Content

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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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