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DUO82065

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Duolink® In Situ Microplate Heat Transfer Block

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About This Item

UNSPSC Code:
41121800
NACRES:
NA.32

product line

Duolink®

technique(s)

proximity ligation assay: suitable

suitability

suitable for brightfield
suitable for fluorescence

storage temp.

20-25°C

General description

Duolink® In Situ Microplate Heat Transfer Block is an aluminium block ideal for keeping an even temperature across the plate during the incubation steps when using Duolink In Situ reagents in multiwell plates. Incubation in the pre-heated Heat Transfer Block increases staining efficiency and reproducibility between different plates, and reduces plate edge effects.

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.

This product can be applied to both the Duolink® In Situ Fluorescence Protocol and the Duolink® In Situ Brightfield Protocol depending on the detection reagents 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.HRP is also available for brightfield detection.
Specificity
Duolink® In Situ Microplate Heat Transfer Block will increase staining efficiency and reproducibility between different plates and reduce edge effects when using Duolink® reagents in microtiter plates.

Keep the Heat Transfer Block preheated at 37°C and keep it in the 37°C incubator throughout the whole Duolink® In Situ protocol. Place the microtiter plate in the Heat Transfer Block during incubation steps.

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.

Linkage
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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

Preparation Note

Storage and Stability: When not in use, store the Heat Transfer Block at room temperature.

Legal Information

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

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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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
Ajay K Yadav et al.
JAMA, 302(3), 276-289 (2009-07-16)
Glioblastomas--uniformly fatal brain tumors--often have both monosomy of chromosome 10 and gains of the epidermal growth factor receptor (EGFR) gene locus on chromosome 7, an association for which the mechanism is poorly understood. To assess whether coselection of EGFR gains
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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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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