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Visualization of RAS/MAPK Signaling In Situ by the Proximity Ligation Assay (PLA).

Methods in molecular biology (Clifton, N.J.) (2016-12-08)
Zijian Tang, Chengkai Dai
ABSTRACT

RAS/MAPK signaling responds to diverse extracellular cues and regulates a wide array of cellular processes. Given its biological importance, abnormalities in RAS/MAPK signaling cascade have been intimately implicated in numerous human diseases, including cancer. Herein, we describe a novel methodology to study activation of this pivotal signaling pathway. The Proximity Ligation Assay (PLA) is employed to monitor kinase-substrate interactions between MEK1 and HSF1, or MEK1 and ERK1 in situ.

MATERIALS
Product Number
Brand
Product Description

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