Duolink® PLA Product Selection Guide

Duolink® PLA Starter Kits

Duolink PLA Starter KitGetting started with Duolink® PLA is simple. The protocols are very similar to that of IF, IHC, and flow cytometry, however Duolink® PLA offers distinct advantages over these and other traditional methods of protein detection.

The Duolink® PLA Starter Kit contains all the necessary reagents you need to perform a fluorescent Duolink® PLA experiment and analyze up to 30 samples. All you have to provide are prepared cells or tissue samples, primary antibodies, and common laboratory equipment.

The Duolink® PLA Starter Kit includes:

Duolink® PLA Probes

Duolink PLA ProbesDuolink® PLA Probes can be purchased separately. They are secondary antibodies conjugated with either the Duolink® PLA PLUS or Duolink® PLA MINUS oligonucleotides. These secondary antibodies are donkey anti-mouse, anti-rabbit, or anti-goat polyclonal antibodies that exhibit minimal cross-reactivity to other species.

There are six options to choose from:

Select those that match the host species of your primary antibodies. Both the Duolink® PLA PLUS probe and Duolink® PLA MINUS probe are required for a successful experiment. In addition, each Duolink® PLA probe kit comes with Blocking Buffer and Antibody Diluent, which have been optimized to work with Duolink® PLA reagents to minimize nonspecific binding of the antibodies and the detection oligonucleotides.

Duolink® Probemaker Kits

For experiments where the Duolink® PLA Probes are not an option because the host species of a primary antibody is different than mouse, rabbit or goat, Duolink® Probemaker kits enable the conjugation of a PLA oligo (PLUS or MINUS) to any antibody. Antibody conjugation with Duolink® Probemaker is quick, easy and requires no clean-up. View the protocol to Create your own PLA® probes for more information.

Duolink® PLA Fluorescent Detection

Duolink PLA Fluorescent detection reagentsDuolink® PLA Fluorescent Detection reagents are available with four different fluorophores: green, orange, red, or far red. Each kit contains the following:

  • 5x Ligation buffer - Contains oligonucleotides that hybridize to the PLA probes and components needed for ligation
  • Ligase enzyme
  • 5x Amplification buffer - Contains components needed for rolling-circle amplification (RCA) and fluor-labeled detection oligonucleotides that hybridize to the RCA product
  • Polymerase

Duolink® PLA Detection Reagents should be selected based on desired fluorescence color and filters available.

Color of Fluorescence Excitation (nm) Emission (nm) Filter (same as for)
Green 495 527 FITC
Orange 554 579 Cyanine 3
Red 594 624 Texas Red
Far Red 644 669 Cyanine 5

Duolink® PLA Brightfield Detection

Duolink® PLA detection is also available for brightfield application. Similar to IHC, Duolink® PLA Brightfield Detection reagents contain oligonucleotides labeled with horseradish peroxidase (HRP) that, in the presence of an HRP substrate, forms an enzyme:substrate precipitate visible using a light microscope. View the Brightfield protocol for more detailed information.

Each Duolink® PLA Brightfield Detection kit contains two packages with the following:

  • Box A (stored at -20°C)
    • 5x Ligation buffer - Contains oligonucleotides that hybridize to the PLA probes and components needed for ligation
    • Ligase enzyme
    • 5x Amplification buffer - Contains components needed for RCA
    • Polymerase
    • 5x Detection buffer - Contains HRP-labeled detection oligonucleotides that hybridize to the RCA product
  • Box B (stored at 4°C)
    • Hydrogen peroxide
    • HRP substrate solutions
    • Nuclear stain

Duolink® PLA Accessories

Additional Duolink® PLA reagents and accessories have been optimized to work with the Duolink® PLA reagents listed above, including:

Product Selection

Determine which products to purchase for your Duolink® PLA experiment depending on your experimental design and application format.  All Duolink® PLA experiments require the use of oligonucleotide-conjugated antibodies, one PLUS and one MINUS. Use the Duolink® PLA Probes if the host species of your primary antibodies are mouse, rabbit, or goat.  If not, Duolink® Probmaker kits for PLUS and MINUS can be used as an alternative. Note: Duolink® Probemaker-made probes can be used in combination with Duolink® PLA Probes as long as there remains one PLUS and one MINUS probe.

For application specific requirements, view the following protocols:


Detection by Fluorescent Microscopy

Required Reagents:

  1. Duolink® PLA Probes (one PLUS and one MINUS from different species, matching the host species of your primary antibodies) OR Duolink® Probmaker PLUS and/or MINUS
  2. Duolink® Fluorescent Detection Reagent (choose from green, orange, red, and far red)
  3. Wash Buffers for Fluorescence
  4. Duolink® Mounting Media with DAPI (for use on slides) OR Nuclear Stain and Anti-fade (for use in microplates)

Optional:

  1. Microplate Heat Transfer Block


Detection by Flow Cytometry

Required Reagents:

  1. Duolink® PLA Probes (one PLUS and one MINUS from different species, matching the host species of your primary antibodies) OR Duolink® Probmaker PLUS and/or MINUS
  2. Duolink® Fluorescent Detection Reagent (choose from green, orange, red, and far red)
  3. Wash Buffers for Fluorescence


Detection by Brightfield Microscopy

Required Reagents:

  1. Duolink® PLA Probes (one PLUS and one MINUS from different species, matching the host species of your primary antibodies) OR Duolink® Probmaker PLUS and/or MINUS
  2. Duolink® Brightfield Detection Reagent
  3. Wash Buffers for Brightfield
  4. Mounting Media for Brightfield OR IHC-compatible mounting media

Optional:

  1. Microplate Heat Transfer Block

Materials

Product No. Description Add to Cart
Starter Kits
DUO92101 Duolink® In Situ Red Starter Kit Mouse/Rabbit
DUO92102 Duolink® In Situ Orange Starter Kit Mouse/Rabbit
DUO92103 Duolink® In Situ Red Starter Kit Mouse/Goat
DUO92104 Duolink® In Situ Orange Starter Kit Mouse/Goat
DUO92105 Duolink® In Situ Red Starter Kit Goat/Rabbit
DUO92106 Duolink® In Situ Orange Starter Kit Goat/Rabbit
Duolink® PLA Accessories
DUO80102 Duolink® In Situ Brightfield Mounting Medium
DUO82040 Duolink® In Situ Mounting Medium with DAPI
DUO82047 Duolink® In Situ Wash Buffer, Brightfield
DUO82049 Duolink® In Situ Wash Buffers, Fluorescence
DUO82064 Duolink® In Situ Microplate Nuclear Stain, Anti-Fade
DUO82065 Duolink® In Situ Microplate Heat Transfer Block
Duolink® PLA Detection Reagents
DUO92007 Duolink® In Situ Detection Reagents Orange
DUO92008 Duolink® In Situ Detection Reagents Red
DUO92012 Duolink® In Situ Detection Reagents Brightfield
DUO92013 Duolink® In Situ Detection Reagents FarRed
DUO92014 Duolink® In Situ Detection Reagents Green
Duolink® PLA Probes
DUO92001 Duolink® In Situ PLA® Probe Anti-Mouse PLUS Affinity purified Donkey anti-Mouse IgG (H+L)
DUO92002 Duolink® In Situ PLA® Probe Anti-Rabbit PLUS Affinity purified Donkey anti-Rabbit IgG (H+L)
DUO92003 Duolink® In Situ PLA® Probe Anti-Goat PLUS Affinity purified Donkey anti-Goat IgG (H+L)
DUO92004 Duolink® In Situ PLA® Probe Anti-Mouse MINUS Affinity purified Donkey anti-Mouse IgG (H+L)
DUO92005 Duolink® In Situ PLA® Probe Anti-Rabbit MINUS Affinity purified Donkey anti-Rabbit IgG (H+L)
DUO92006 Duolink® In Situ PLA® Probe Anti-Goat MINUS Affinity purified Donkey anti-Goat IgG (H+L)
DUO92009 Duolink® In Situ Probemaker PLUS
DUO92010 Duolink® In Situ Probemaker MINUS

back to top