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DUO82049

Duolink® In Situ Wash Buffers, Fluorescence

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4 L
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$86.90
20 L
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$334.00

About This Item

NACRES:
NA.32
UNSPSC Code:
12161703

$86.90

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material

packing (powdered buffer pouches)

Quality Segment

200

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for fluorescence

storage temp.

20-25°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.

Use the Duolink® In Situ Fluorescence Protocol for 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. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), 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
Duolink® In Situ fluorescence applications use two wash buffers. Wash Buffer A is used after the PLA Probe incubation step and Wash Buffer B is used after incubation with the amplification reagents. See datasheet for more information.

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.

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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).

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

Legal Information

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

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technique(s)

proximity ligation assay: suitable

technique(s)

proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

proximity ligation assay: suitable

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

-

product line

Duolink®

product line

Duolink®

product line

Duolink®

product line

Duolink®

storage temp.

20-25°C

storage temp.

20-25°C

storage temp.

−20°C

storage temp.

−20°C

suitability

suitable for fluorescence

suitability

suitable for brightfield

suitability

suitable for brightfield, suitable for fluorescence

suitability

suitable for brightfield

material

packing (powdered buffer pouches)

material

packing (powdered buffer pouches)

material

-

material

-

Storage Class

10 - Combustible liquids

wgk

WGK 3

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Questions

  1. Which fluorophore used in PLA assay (green, red or infrared) is suitable to use for STED microscopy?

    1 answer

    1. This product is a fluorescence wash buffer used across PLA fluorescence assays. In general, Texas Red and FITC have been frequently cited for use in STED microcopy. In addition, the exact nature of the DuoLink PLA fluorochromes are considered proprietary and may not be disclosed. The stability of these fluorochromes in STED experiments has not been determined. Please refer to the image below listing the emission/excitation as well as the recommended filter.

      For STED, it is recommended a far-red staining. STED literature and a recent multi color FLIM STED study used far red dyes (ATTO 647N, Abberior STAR 635P, CF680R, Alexa/CF594 etc.) and recommends using a single 775 nm pulsed depletion laser for multi color STED.
      https://pmc.ncbi.nlm.nih.gov/articles/PMC9388687/

      Another study of interest, used different PLA approaches with STED microscopy including the Duolink® In Situ Detection Reagents Red.
      https://pmc.ncbi.nlm.nih.gov/articles/PMC11607433/

      Note that verification of the dye photostability/lifetime under specific STED conditions (STED power, depletion wavelength, conjugation partner) is strongly recommended as the lifetime and bleaching behavior can change.

      Helpful?

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