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.SpecificityThis 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 noteIn 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 proteinLet us do the work for you, learn more about our Custom Service Program to accelerate your Duolink^® projectsView full Duolink^® product list

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.

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

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.

Legal Information

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

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

Other Notes

Other Duolink^® reagents needed (30RXN size):Mouse/Rabbit Duolink^® PLA Starter Kit: Red Starter Kit (DUO92101) OR Orange Starter Kit (DUO92102)ORPLA 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).