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In Situ Proximity Ligation Assay to Detect the Interaction Between Plant Transcription Factors and Other Regulatory Proteins.

Methods in molecular biology (Clifton, N.J.) (2018-07-26)
Yifeng Xu, Eng-Seng Gan, Toshiro Ito
ABSTRACT

In plants, a lot of transcription factors fulfill their roles in gene regulation through the interaction with other regulatory proteins and co-factors. Thus, confirmation of protein-protein interaction is key to understand the precise function of transcription factors. Many methods have been developed to investigate the protein-protein interaction in vivo and in vitro. In situ Proximity Ligation Assay (PLA) is an innovative method to test protein-protein interaction in your tissues or cells of interest in vivo. Furthermore, this method allows us to detect transient interaction and low-abundance protein interaction with single molecule resolution. In this chapter, we describe a detailed protocol for the study of interaction between plant transcription factors and other regulatory proteins, in the scale of single nuclei of plant organ, tissues and cells.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Duolink® In Situ Red Starter Kit Mouse/Rabbit
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Duolink® In Situ Detection Reagents Red
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Duolink® In Situ PLA® Probe Anti-Mouse MINUS, Affinity purified Donkey anti-Mouse IgG (H+L)
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Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, Affinity purified Donkey anti-Rabbit IgG (H+L)
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Duolink® In Situ Orange Starter Kit Mouse/Rabbit
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Duolink® In Situ PLA® Probe Anti-Mouse PLUS
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Duolink® In Situ PLA® Probe Anti-Rabbit MINUS
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Duolink® In Situ Detection Reagents Orange
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Duolink® In Situ Wash Buffers, Fluorescence
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Duolink® In Situ Detection Reagents Green
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Duolink® In Situ Detection Reagents FarRed
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Duolink® In Situ Mounting Medium with DAPI
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Duolink® In Situ Probemaker MINUS
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Duolink® In Situ Probemaker PLUS
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Duolink® In Situ Detection Reagents Brightfield
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Duolink® flowPLA Detection Kit - Green, Duolink® PLA kit for Flow Cytometry with Green Detection
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Duolink® In Situ PLA® Probe Anti-Goat PLUS
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Duolink® flowPLA Detection Kit - FarRed, Duolink® PLA kit for Flow Cytometry with FarRed Detection
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Duolink® PLA Control Kit - PPI, Control Kit to reliably detect Protein-Protein Interactions (PPI) with Duolink® PLA
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Duolink® In Situ PLA® Probe Anti-Goat MINUS, Affinity purified Donkey anti-Goat IgG (H+L)
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Duolink® In Situ Wash Buffer, Brightfield
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Duolink® flowPLA Detection Kit - Red, DUOLINK® : PLA kit for Flow Cytometry with Red Detection
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Duolink® In Situ Red Starter Kit Mouse/Goat
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Duolink® In Situ Red Starter Kit Goat/Rabbit
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Duolink® In Situ Microplate Nuclear Stain, Anti-Fade
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Duolink® flowPLA Detection Kit - Orange, Duolink® PLA kit for Flow Cytometry with Orange Detection
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Duolink® In Situ Orange Starter Kit Goat/Rabbit
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Duolink® In Situ Orange Starter Kit Mouse/Goat
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Duolink® In Situ Microplate Heat Transfer Block