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DUO92014

Sigma-Aldrich

Duolink® In Situ Detection Reagents Green

NACRES:
NA.32

Quality Level

product line

Duolink®

technique(s)

proximity ligation assay: suitable

fluorescence

λex 495 nm; λem 527 nm (green) (FITC (Cyanine 2), Zeiss Filter set 38)

suitability

suitable for fluorescence

shipped in

dry ice

storage temp.

−20°C

General description

Duolink® In Situ Detection Reagents Green contains all the necessary Duolink In situ reagents to perform the amplification and detection of bound PLA® probes. The detection probes contain a fluorophore (lex = 495 nm and lem = 527 nm), which may be visualized using the same filter as Cy®2 or FITC. Experiments conducted using Duolink In situ reagents can detect and visualize protein interactions, protein expression levels and post translational modifications at the single molecule level in fixed cells and tissue samples.

Specificity

Green fluorescence detection reagents are often used with FITC filter.

Application

Duolink® In Situ Detection Reagents has been used in the proximity ligation assay of:
  • vasopressin and gonadotropin-releasing hormone (GnRH) from frozen rat brain sections
  • human embryonic kidney 293 cells (HEK)
  • pituitary tissues

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.

Features and Benefits

  • No overexpression or genetic manipulation reNo 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

Components

  • 5x Ligation - Contains oligonucleotides that hybridize to the PLA probes and all components needed for ligation except the Ligase
  • 1x Ligase (1 unit/μL)
  • 1x Polymerase (10 units/μL)
  • 5x Amplification Green - 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.
See datasheet for more information.

Not included in Detection kit:

Primary antibodies, PLA probes, wash buffers, mounting medium

Preparation Note

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.

Storage and Stability

Store the components at –20 °C. The enzymes should be kept cold (–20 °C) at all times, use a freezing block when removing them from the freezer.

Other Notes

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

View full Duolink® product list

Legal Information

Cy is a registered trademark of Cytiva
Duolink is a registered trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Certificate of Analysis

Certificate of Origin

Protein Phosphatase 1c Associated with the Cardiac Sodium Calcium Exchanger1 Regulates its Activity by Dephosphorylating Serine 68 Phosphorylated Phospholemman.
Hafver T L, et al.
The Journal of Biological Chemistry, M115-M115 (2015)
Longze Sha et al.
The Journal of experimental medicine, 214(2), 547-563 (2016-12-29)
The glutamate transporter GLT-1 is critical for the maintenance of low interstitial glutamate concentrations. Loss of GLT-1 is commonly observed in neurological disorders, including temporal lobe epilepsy (TLE). Despite the hypothesis that targeting the mechanisms of GLT-1 deficiency may be
Xiaofan Li et al.
PLoS pathogens, 13(3), e1006249-e1006249 (2017-03-02)
Trials to reintroduce chloroquine into regions of Africa where P. falciparum has regained susceptibility to chloroquine are underway. However, there are long-standing concerns about whether chloroquine increases lytic-replication of Epstein-Barr virus (EBV), thereby contributing to the development of endemic Burkitt
Xiaofan Li et al.
Journal of virology, 93(17) (2019-06-14)
Herpesviruses are ubiquitous, and infection by some, like Epstein-Barr virus (EBV), is nearly universal. To persist, EBV must periodically switch from a latent to a replicative/lytic phase. This productive phase is responsible for most herpesvirus-associated diseases. EBV encodes a latency-to-lytic
Birgitte B Olsen et al.
BMC molecular biology, 13, 7-7 (2012-03-13)
The DNA-dependent protein kinase (DNA-PK) is a nuclear complex composed of a large catalytic subunit (DNA-PKcs) and a heterodimeric DNA-targeting subunit Ku. DNA-PK is a major component of the non-homologous end-joining (NHEJ) repair mechanism, which is activated in the presence

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