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DUO92101

Duolink® In Situ Red Starter Kit Mouse/Rabbit

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About This Item

UNSPSC Code:
41116158
NACRES:
NA.32

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product line

Duolink®

technique(s)

proximity ligation assay: suitable

fluorescence

λex 594 nm; λem 624 nm (Texas Red®; Zeiss Filter set 31)

suitability

suitable for fluorescence

storage temp.

−20°C

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This Item
DUO92008DUO92013DUO92007
Duolink® In Situ Red Starter Kit Mouse/Rabbit

Sigma-Aldrich

DUO92101

Duolink® In Situ Red Starter Kit Mouse/Rabbit

Duolink® In Situ Detection Reagents Red

Sigma-Aldrich

DUO92008

Duolink® In Situ Detection Reagents Red

Duolink® In Situ Detection Reagents FarRed

Sigma-Aldrich

DUO92013

Duolink® In Situ Detection Reagents FarRed

Duolink® In Situ Detection Reagents Orange

Sigma-Aldrich

DUO92007

Duolink® In Situ Detection Reagents Orange

technique(s)

proximity ligation assay: suitable

technique(s)

proximity ligation assay: suitable

technique(s)

proximity ligation assay: suitable

technique(s)

proximity ligation assay: suitable

suitability

suitable for fluorescence

suitability

suitable for fluorescence

suitability

suitable for fluorescence

suitability

suitable for fluorescence

fluorescence

λex 594 nm; λem 624 nm (Texas Red®; Zeiss Filter set 31)

fluorescence

λex 594 nm; λem 624 nm (Texas Red®, Zeiss Filter set 31)

fluorescence

λex 644 nm; λem 669 nm (Cyanine 5, Zeiss Filter set 50)

fluorescence

λex 554 nm; λem 576 nm (Cyanine 3; Zeiss Filter set 20)

product line

Duolink®

product line

Duolink®

product line

Duolink®

product line

Duolink®

Quality Level

100

Quality Level

200

Quality Level

200

Quality Level

200

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Application

Duolink®proximity ligation assay(PLA®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

Follow the Duolink® In Situ Fluorescence Protocol to use this product. A set of short instructionsis also available.

Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes.This starter kit supplies all other necessary reagents for 30 Duolink® PLA reactions, which include a pair of PLA probes (Anti-Rabbit PLUS and Anti-Mouse MINUS), red detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink® PLA probes. Analysis is carried out using standard immunofluorescence assay equipment.
Specificity
The Duolink® In Situ Red Starter Kit Mouse/Rabbit requires one primary antibody from mouse and one primary antibody from rabbit. Red fluorescence detection reagents are often used with Texas Red® filter.

Application Note
Two primary antibodies raised in different species are needed. Test your primary antibodies (IgG-class, mono- or polyclonal) in a standard immunofluorescence (IF), immunohistochemistry (IHC) or immunocytochemistry (ICC) assay to determine the optimal fixation, blocking, and titer conditions. Duolink® PLA in situ reagents are suitable for use on fixed cells, cytospin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

View full Duolink® product list
Duolink® in situ red starter kit has been used in proximity ligation assay.[1][2]

Features and Benefits

  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Single molecule sensitivity due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Quicker and simpler than FRET
  • Increased accuracy compared to co-IP
  • Publication-ready results

Other Notes

2024 CiteAb Award Winner for Supplier Succeeding in Parkinson′s Research

Legal Information

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
Texas Red is a registered trademark of Life Technologies

Kit Components Also Available Separately

Product No.
Description
SDS
  • DUO92002Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, Affinity purified Donkey anti-Rabbit IgG (H+L)SDS
  • DUO92004Duolink® In Situ PLA® Probe Anti-Mouse MINUS, Affinity purified Donkey anti-Mouse IgG (H+L)SDS
  • DUO92008Duolink® In Situ Detection Reagents RedSDS
  • DUO82049Duolink® In Situ Wash Buffers, FluorescenceSDS
  • DUO82040Duolink® In Situ Mounting Medium with DAPISDS

Signal Word

Danger

Hazard Statements

H290,H317,H334,H411

Hazard Classifications

Aquatic Chronic 2 - Met. Corr. 1 - Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

8A - Combustible corrosive hazardous materials

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Certificates of Analysis (COA)

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New protein?protein interactions of mitochondrial connexin 43 in mouse heart.
Denuc A, et al.
Journal of Cellular and Molecular Medicine, 20(5), 794-803 (2016)
JNK associated leucine zipper protein functions as a docking platform for Polo like kinase 1 and regulation of the associating transcription factor Forkhead box protein K1.
Ramkumar P, et al.
The Journal of Biological Chemistry (2015)
Targeting CPT1A-mediated fatty acid oxidation sensitizes nasopharyngeal carcinoma to radiation therapy.
Tan Z, et al.
Theranostics, 8(9), 2329-2329 (2018)
Roberto Di Maio et al.
Science translational medicine, 10(451) (2018-07-27)
Missense mutations in leucine-rich repeat kinase 2 (LRRK2) cause familial Parkinson's disease (PD). However, a potential role of wild-type LRRK2 in idiopathic PD (iPD) remains unclear. Here, we developed proximity ligation assays to assess Ser1292 phosphorylation of LRRK2 and, separately
Ivan Duran et al.
Human molecular genetics, 24(7), 1918-1928 (2014-12-17)
Osteogenesis imperfecta (OI) is a genetic disorder that results in low bone mineral density and brittle bones. Most cases result from dominant mutations in the type I procollagen genes, but mutations in a growing number of genes have been identified
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Duolink®邻位连接技术用于研究神经元相互作用，促进神经科学研究。

Duolink® PLA故障排除指南

包括建议和技巧、常见问题解答和基本故障排除的支持信息。

Duolink® PLA荧光实验方案

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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.

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Protocols

Duolink® In Situ Short Instructions - Fluorescence

This page details the Duolink® In Situ Short Protocol for fluorescence detection

Duolink® PLA Fluorescence Protocol

This protocol describes how to perform immunofluorescent detection of proteins in cells and tissue.

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蛋白质与核酸的相互作用

蛋白质与核酸相互作用试剂，以及用于研究蛋白质-RNA、蛋白质-DNA、蛋白质-蛋白质相互作用和相关应用的资源。

Duolink® PLA邻位连接技术的应用

利用邻位连接技术对蛋白质互作、翻译后修饰和低表达蛋白进行检测、定量和成像应用。

Duolink® PLA Applications

Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay

Questions

1–8 of 8 Questions  

  1. I want to use Duolink™ in Multiwell Plates: is the Duolink® In Situ Microplate Nuclear Stain, Anti-Fade included in the Duolink In Situ starter kit?

    1 answer

    1. Product: DUO92101 is our Duolink® In Situ Red Starter Kit Mouse/Rabbit.

      This kit contains the following components:

      Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, Affinity purified Donkey anti-Rabbit IgG (H+L) (DUO92002)
      Duolink® In Situ PLA® Probe Anti-Mouse MINUS, Affinity purified Donkey anti-Mouse IgG (H+L) (DUO92004)

      These components are each comprised of the following:
      • 5x PLA Probe Anti-Rabbit PLUS - Donkey anti-rabbit secondary antibody conjugated to oligonucleotide PLUS
      • 1x Blocking Solution - Reagent for blocking of the sample
      • 1x Antibody Diluent - For dilution of PLA probes and primary antibodies

      Duolink® In Situ Detection Reagents Red (DUO92008)

      This component is comprised of the following:
      • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
      • 1x Ligase (1 unit/?L)
      • 1x Polymerase (10 units/?L)
      • 5x Amplification Red - Contains all components needed for Rolling Circle Amplification (RCA) except the Polymerase. It also contains oligonucleotide probes labeled with a fluorophore that hybridize to the RCA product.

      Duolink® In Situ Wash Buffers, Fluorescence (DUO82049)
      Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents.

      Duolink® In Situ Mounting Medium with DAPI (DUO82040)
      Duolink® In Situ Mounting Medium with DAPI is ideal for nuclear staining and preserving signals generated with the Duolink® In Situ Detection Reagents for fluorescence microscopy. Note: Counterstaining with Cy®2 is not recommended.

      The Duolink® In Situ Microplate Nuclear Stain, Anti-Fade is sold separately.

      Helpful?

  2. Which fixation method (formalin, PFA, ethanol, aceton/methanol) do you recommend to use for fresh frozen tissue slides before applying the PLA?

    1 answer

    1. The choice of fixation techniques varies between sample types. Academic literature should be consulted to determine the proven methods of fixation for the specific frozen tissue types used in IHC and PLA experiments to best address this question.

      Helpful?

  3. Does this kit contain antibody diluent and blocking solution? Or do you have to buy them separately?

    1 answer

    1. The kit contains antibody 1X diluent and 1X blocking solution, which can be found in the component DUO92002 and DUO92004.

      Helpful?

  4. Hello, Is this kit also suitable for flow cytometry-based analysis of the data?

    1 answer

    1. This kit is NOT suitable for flow cytometry. Please see our Duolink® flowPLA Mouse/Rabbit Starter Kits for flow cytometry: DUO94102 (Green), DUO94104 (Far Red), & DUO94105 (Violet).

      Helpful?

  5. Can the Duolink kits be used with a Mouse IgM antibody?

    1 answer

    1. Duolink kits may be compatible with a Mouse IgM antibody, provided that the antibody contains the light chain. The Anti-Mouse MINUS probe specifically interacts with mouse IgG (H+L) and the light chains associated with IgM. Therefore, if the Mouse IgM antibody includes the light chain, it should be suitable for use with the Duolink kits.

      Helpful?

  6. How many reactions can be done with this kit?

    1 answer

    1. The Duolink™ starter kit contains all the components necessary to analyze up to 30 reactions in one single order. Please see the link below to review the full protocol:
      https://www.sigmaaldrich.com/US/en/technical-documents/protocol/protein-biology/protein-and-nucleic-acid-interactions/duolink-fluorescence-user-manual#DuolinkPLAFluorescenceProtocol

      Helpful?

  7. I want to study the interaction of two cell surface proteins using PLA kit . My primaries are mouse and rabbit. Does the kit have the two secondary and also all the components that i would require to carry out the assay. Can you advice?

    1 answer

    1. The Duolink® In Situ Red Starter Kit Mouse/Rabbit - DUO92101 contains the Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, affinity purified Donkey anti-Rabbit IgG, and Duolink® In Situ PLA® Probe Anti-Mouse MINUS, affinity purified Donkey anti-Mouse IgG (H+L) secondary antibodies and everything needed to conduct the PLA assay.

      Helpful?

  8. What is the expiry date of the kit contents?

    1 answer

    1. This product is not assigned an expiration date or recommended retest date. Products with no expiration date or recommended retest date should be routinely inspected by customers to ensure they perform as expected. These products are also subject to a one-year warranty from the date of shipment.

      For more information, you may access the "Product Dating Information" document under "ADDITIONAL USEFUL DOCUMENTS ABOUT OUR PRODUCTS" at the bottom of the Quality Services page with this link: https://www.sigmaaldrich.com/life-science/quality-and-regulatory-management/quality-services.

      Helpful?

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