DUO92202

Sigma-Aldrich

Duolink® PLA Control Kit - PPI

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

product line

Duolink®

application(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

    RIDADR

    NONH for all modes of transport

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