DUO92007
Duolink® In Situ Detection Reagents Orange
Synonim(y):
in situ Proximity Ligation Assay reagent, Protein Protein Interaction Assay reagent
About This Item
linia produktu
Duolink®
Poziom jakości
metody
proximity ligation assay: suitable
fluorescencja
λex 554 nm; λem 576 nm (Cyanine 3; Zeiss Filter set 20)
przydatność
suitable for fluorescence
Warunki transportu
dry ice
temp. przechowywania
−20°C
Powiązane kategorie
Zastosowanie
Follow the Duolink® In Situ Fluorescence Protocolto 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.
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.
Orange fluorescence detection reagents are often used with Cyanine 3 filter.
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.
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Cechy i korzyści
- 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
Komponenty
- 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 Orange - 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.
Not included in Detection kit:
Primary antibodies, PLA probes, wash buffers, mounting medium
Informacje prawne
Kod klasy składowania
10 - Combustible liquids
Klasa zagrożenia wodnego (WGK)
WGK 2
Certyfikaty analizy (CoA)
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Produkty
Protocol for immunofluorescent detection of proteins in cells and tissue
Dowiedz się, jak działa technologia Proximity Ligation Assay i jak zestaw kontrolny interakcji białko-białko może potwierdzić wykrywanie in situ dimeryzacji EGFR-HER2 indukowanej przez EGF.
Find Duolink references based on the type of method used, post translational modification detected, and research focus.
Things to consider for preparation, setup and execution of the Duolink® assay protocol
Protokoły
Niniejszy protokół opisuje sposób przeprowadzania immunofluorescencyjnego wykrywania białek w komórkach i tkankach.
This page details the Duolink® In Situ Short Protocol for fluorescence detection
Nasz zespół naukowców ma doświadczenie we wszystkich obszarach badań, w tym w naukach przyrodniczych, materiałoznawstwie, syntezie chemicznej, chromatografii, analityce i wielu innych dziedzinach.
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