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DUO92202

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Duolink® PLA Control Kit - PPI

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

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Synonym(s):
in situ Proximity Ligation Assay Control Kit, Protein Protein Interaction Assay Control Kit

product line

Duolink®

technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

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DUO96020DUO96030DUO96010
technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, multiplexing: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, multiplexing: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, multiplexing: suitable, proximity ligation assay: suitable

suitability

suitable for fluorescence

suitability

suitable for fluorescence

suitability

suitable for fluorescence

suitability

suitable for fluorescence

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

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

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)

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 Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

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.

Storage Class Code

11 - Combustible Solids


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Franziska Wetzel et al.
Cellular and molecular life sciences : CMLS, 74(2), 373-392 (2016-09-09)
The zonula occludens (ZO)-2 protein links tight junctional transmembrane proteins to the actin cytoskeleton and associates with splicing and transcription factors in the nucleus. Multiple posttranslational modifications control the intracellular distribution of ZO-2. Here, we report that ZO-2 is a
Valerie Le Sage et al.
Virology, 502, 73-83 (2016-12-26)
Stress granules (SGs) are dynamic cytoplasmic aggregates of translationally silenced mRNAs that assemble in response to environmental stress. SGs appear to play an important role in antiviral innate immunity and many viruses have evolved to block or subvert SGs components
Teng-Jian Zhou et al.
Theranostics, 7(5), 1389-1406 (2017-04-25)
Cancer stem cells (CSCs) are a small subset of malignant cells, possessing stemness, with strong tumorigenic capability, conferring resistance to therapy and leading to the relapse of nasopharyngeal carcinoma (NPC). Our previous study suggested that cyclooxygenase-2 (COX-2) would be a
Fredrick J Rosario et al.
Scientific reports, 7(1), 3982-3982 (2017-06-24)
Maternal folate deficiency is linked to restricted fetal growth, however the underlying mechanisms remain to be established. Here we tested the hypothesis that mTOR functions as a folate sensor in vivo in mice and that maternal folate deficiency inhibits placental
Timothy L Scott et al.
Molecular carcinogenesis, 56(2), 325-336 (2016-05-06)
Apurinic/apyrimidinic endonuclease 1 (APE1) is an essential protein crucial for repair of oxidized DNA damage not only in genomic DNA but also in mitochondrial DNA. Parkin, a tumor suppressor and Parkinson's disease (PD) associated gene, is an E3 ubiquitin ligase

Articles

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.

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

Protocols

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.

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