DUO92005
Duolink® In Situ PLA® Probe Anti-Rabbit MINUS
biological source
donkey (polyclonal)
Quality Level
antibody form
affinity purified immunoglobulin (secondary antibody)
antibody product type
primary antibodies
product line
Duolink®
species reactivity
rabbit
technique(s)
immunofluorescence: suitable
proximity ligation assay: suitable
suitability
suitable for brightfield
suitable for fluorescence
shipped in
wet ice
storage temp.
2-8°C
Related Categories
Application
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.
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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.
PLA probe anti-Rabbit reacts with whole molecule rabbit IgG and the light chains of other rabbit immunoglobulin?s. The PLA probe anti-Rabbit has minimal cross reactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, human, mouse, rat, and sheep serum proteins. A PLUS probe of a different species must be used simultaneously with this product. See our Product Selection Guide 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® PLA 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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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
- 5x PLA Probe Anti-Rabbit MINUS - Donkey anti-rabbit secondary antibody conjugated to oligonucleotide MINUS
- 1x Blocking Solution - Reagent for blocking of the sample
- 1x Antibody Diluent - For dilution of PLA probes and primary antibodies
Legal Information
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
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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Example:
Additional examples:
705578-5MG-PW
PL860-CGA/SHF-1EA
MMYOMAG-74K-13
1000309185
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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').
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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.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
What epitope on the antibody is recognized by the Duolink PLA Probes?
The Duolink PLA Probes offered are donkey polyclonal antibodies directed to the whole molecule (heavy and light chains). They are affinity purified. The same polyclonal antibody is conjugated to the Plus and Minus Oligos. The Plus and Minus probes can be used together (to detect a single antibody) since they can recognize multiple sites. The anti-mouse IgG PLA labeled antibodies may cross-react with rat but this is not consistent. Do not use this product with rat antibodies. It is best to label anti-rat secondary antibodies with the Duolink Probemaker kit.The anti-goat IgG PLA labeled antibodies have cross-reactivity with sheep IgG and can be recommended for this application.
Has Duolink been used for detection of changes in protein expression?
The following reference cites the use of Duolink to detect changes in protein expression in Human. Raizer et al., Cancer 2010;116:5297-305
How many reactions can be performed with the Duolink® In Situ Starter Kit?
All of the starter kits for the Duolink product line will perform 30 reactions using 40 ul per reaction. DUO92101 - Duolink® In Situ Red Starter Kit Mouse/Rabbit DUO92102 - Duolink® In Situ Orange Starter Kit Mouse/Rabbit DUO92103 - Duolink® In Situ Red Starter Kit Mouse/Goat DUO92104 - Duolink® In Situ Orange Starter Kit Mouse/Goat DUO92105 - Duolink® In Situ Red Starter Kit Goat/Rabbit DUO92106 - Duolink® In Situ Orange Starter Kit Goat/Rabbit
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Articles
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
This protocol describes the use of Duolink® PLA reagents for the brightfield detection, visualization, and quantification of individual proteins, protein modifications, and protein interactions in tissue and cell samples.
Protocol for use of Duolink® PLA reagents for the detection of individual proteins, protein modifications, and protein-protein interactions within cell populations by flow cytometry.
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