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DUO82064

Sigma-Aldrich

Duolink® In Situ Microplate Nuclear Stain, Anti-Fade

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Synonym(s):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
NACRES:
NA.32

product line

Duolink®

Quality Level

technique(s)

proximity ligation assay: suitable

fluorescence

λex 360 nm; λem 460 nm

suitability

suitable for fluorescence-detection automated sequencing
suitable for microtiter plates

shipped in

dry ice

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.

Use the Multiwell Plates modifications to the Duolink® In Situ Fluorescence Protocol to run an experiment with this product. A set of short instructions can also be used.

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 Microplate Nuclear Stain and Anti-Fade are intended to be used after staining cells with Duolink® In Situ in microtiter plates. See the 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® 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

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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

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Aquatic Chronic 3 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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705578-5MG-PW

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Thomas W Bonagura et al.
Endocrinology, 153(6), 2897-2906 (2012-04-13)
We previously showed that advancing the increase in estradiol levels from the second to the first third of baboon pregnancy suppressed placental extravillous trophoblast (EVT) invasion and remodeling of the uterine spiral arteries. Cell culture studies show that vascular endothelial
Jaclyn J Renfrow et al.
Neuro-oncology, 13(8), 880-885 (2011-07-30)
We present a novel methodology combining traditional fluorescent in situ hybridization with an in situ protein detection technology called proximity ligation assay. This method has potential to perform a detailed analysis of the relationship between gene status and corresponding protein
Jeffrey J Raizer et al.
Cancer, 116(22), 5297-5305 (2010-07-29)
The authors evaluated a 3-week schedule of bevacizumab in patients with recurrent high-grade glioma (HGG). Patients received bevacizumab 15 mg/kg every 3 weeks and were evaluated every 6 weeks until tumor progression. Tissue correlates were used to quantify tumor content
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
Charles Lu et al.
PloS one, 7(4), e34833-e34833 (2012-05-05)
Tumor suppressor gene TUSC2/FUS1 (TUSC2) is frequently inactivated early in lung cancer development. TUSC2 mediates apoptosis in cancer cells but not normal cells by upregulation of the intrinsic apoptotic pathway. No drug strategies currently exist targeting loss-of-function genetic abnormalities. We

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Things to consider for preparation, setup and execution of the Duolink® assay protocol

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

Protocols

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

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