Recommended Products
product line
Duolink®
technique(s)
immunofluorescence: suitable
proximity ligation assay: suitable
suitability
suitable for brightfield
suitable for fluorescence
shipped in
wet ice
storage temp.
−20°C
Related Categories
Application
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Specificity
Duolink® In Situ Probemaker extends your possibilities even further by allowing you to create your own PLA probes to meet specific assay requirements.
- Study homodimers
- Study protein-protein interactions with two primary antibodies derived from the same species
- Study protein-protein interactions with primary antibodies from any species
Using Duolink® In Situ Probemaker PLUS and MINUS, you simply conjugate the PLUS and MINUS oligo arms directly to your antibodies in a quick and convenient procedure. As well as providing unique capabilities, this also means there is no longer any limitation as to which species your primary antibodies have to be derived from in order perform a Duolink® In Situ assay.
Application Note
The antibody to be conjugated must have a concentration of 1 mg/ml. Do not use volumes larger than 20 ml for conjugation. The antibody must be in an amine free buffer, ideally PBS. The buffer should be carrier free but may contain up to 0.1% BSA, 5% trehalose, and 0.02% sodium azide.
- Study homodimers by using one monoclonal antibody split into two halves. Label one with the PLUS oligo and the other with the MINUS oligo.
- Use antibodies from same species: study protein-protein interactions, protein modifications, or single proteins, using two primary antibodies derived from the same species. Label one of the antibodies with the PLUS oligo and the other with the MINUS oligo
- Use antibodies from any species: study protein-protein interactions, protein modifications, or single proteins, using one or both primary antibodies derived from species other than mouse, rabbit or goat. Label one of your secondary antibodies with the PLUS oligo and the other with the MINUS oligo. Or, combine one labeled secondary antibody with a standard secondary Duolink®In Situ PLA probe.
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
Components
- Duolink® In Situ oligonucleotide PLUS – one vial with lyophilized activated PLUS oligonucleotide for one conjugation of 20 μg antibody
- Conjugation buffer – buffer for conjugation reaction
- Stop reagent – stops conjugation reaction
- Storage solution – buffer for preserving prepared PLA probe (conjugated antibody)
- 20x Assay reagent – reagent to be added to experimenter optimized antibody diluent
- Blocking solution – blocks sample prior to staining with Duolink® In Situ
- PLA probe diluent – buffer for diluting PLA probe (conjugated antibody) to final assay concentration
Other Notes
Follow the Duolink®In Situ Probemaker Protocol to use the Duolink®In Situ Probemaker PLUS and MINUS.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.For more information, see Create Your Own PLA® Probes.
Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.
Legal Information
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Aquatic Chronic 2 - Skin Sens. 1
Storage Class Code
10 - Combustible liquids
Regulatory Information
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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How to Find the Product Number
Product numbers are combined with Pack Sizes/Quantity when displayed on the website (example: T1503-25G). Please make sure you enter ONLY the product number in the Product Number field (example: T1503).
Example:
Additional examples:
705578-5MG-PW
PL860-CGA/SHF-1EA
MMYOMAG-74K-13
1000309185
enter as 1.000309185)
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How to Find a Lot/Batch Number for COA
Lot and Batch Numbers can be found on a product's label following the words 'Lot' or 'Batch'.
Aldrich Products
For a lot number such as TO09019TO, enter it as 09019TO (without the first two letters 'TO').
For a lot number with a filling-code such as 05427ES-021, enter it as 05427ES (without the filling-code '-021').
For a lot number with a filling-code such as STBB0728K9, enter it as STBB0728 without the filling-code 'K9'.
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In some cases, a COA may not be available online. If your search was unable to find the COA you can request one.
Articles
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
Support information including tips and tricks, frequently asked questions, and basic troubleshooting.
Protocols
Duolink® kits use in situ PLA®, a proximity ligation assay technology, to accurately and objectively quantify individual proteins, and their interactions and modifications in unmodified cells and tissue.
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
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