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DUO82064

Sigma-Aldrich

Duolink® In Situ Microplate Nuclear Stain, Anti-Fade

Synonym(s):

in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.32

product line

Duolink®

Quality Level

200

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
GHS07

Signal Word

Warning

Hazard Statements

H317,H412

Hazard Classifications

Aquatic Chronic 3 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Lydie Couturier et al.
Nature cell biology, 14(2), 131-139 (2012-01-24)
Cell-fate diversity can be generated by the unequal segregation of the Notch regulator Numb at mitosis in both vertebrates and invertebrates. Whereas the mechanisms underlying unequal inheritance of Numb are understood, how Numb antagonizes Notch has remained unsolved. Live imaging
Ping-Chih Ho et al.
Nature immunology, 13(4), 379-386 (2012-03-06)
Tolerance to endotoxins that is triggered by prior exposure to Toll-like receptor (TLR) ligands provides a mechanism with which to dampen inflammatory cytokines. The receptor-interacting protein RIP140 interacts with the transcription factor NF-κB to regulate the expression of genes encoding
Yukiko Hasumi et al.
Proceedings of the National Academy of Sciences of the United States of America, 106(44), 18722-18727 (2009-10-24)
Germline mutations in the BHD/FLCN tumor suppressor gene predispose patients to develop renal tumors in the hamartoma syndrome, Birt-Hogg-Dubé (BHD). BHD encodes folliculin, a protein with unknown function that may interact with the energy- and nutrient-sensing AMPK-mTOR signaling pathways. To
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
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
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