Duolink® PLA Probe Mouse Anti-PhosThr MINUS from mouse，50MIG，DUO87014-50UG，Sigma

General description

Anti-phosphothreonine antibody, clone 20H6.1 with the Duolink^® PLA^® MINUS oligonucleotide. This antibody detects threonine phosphorylated proteins in all species. Protein target specificity can be obtained by pairing with an antibody specific to a protein of interest in a Duolink^® PLA^® experiment. This product must be used in combination with a PLUS PLA probe of a different species or another directly-conjugated primary antibody with a PLUS PLA oligo. The Anti-mouse PLA^® probe MINUS (DUO92001) is not necessary when using this product.

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.

Application Note
Protein phosphorylation is considered as one of the key steps in signal transduction and regulation of enzymatic activity. Anti-phosThr antibodies are commonly used in western blots after the targeted proteins have been immunoprecipitated to measure the Threonine phosphorylation of the protein of interest. This process is time-consuming and cannot provide subcellular localization.

Anti-phosThr PLA probes are used to detect the phosphorylation of threonine residues in proteins. Protein target specificity can be obtained by pairing with an antibody specific to a protein of interest in a Duolink^® PLA^® experiment. This product is particularly useful when site-specific anti-phosphorylation antibodies are not available. A PLUS PLA probe of a different species or another directly-conjugated primary antibody with a PLUS PLA oligo must be used in combination with this product. The Anti-mouse PLA probe MINUS (DUO92001) is not necessary when using this product. See our Product Selection Guide for more information.

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Biochem/physiol Actions

This directly-conjugated primary antibody PLA probe recognizes threonine-phosphorylated proteins from all species.

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