Duolink(TM) PLA Control Kit - PPI

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

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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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.

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)

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

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

Disclaimer

RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.