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HCR™ IF + HCR™ RNA-FISH Protocols

Simultaneous multiplexed, quantitative, high-resolution protein and RNA imaging 

HCR™ IF probes, amplifiers, and buffers and HCR™ RNA-FISH probe sets, amplifiers, and buffers enable a unified approach to multiplexed quantitative high-resolution immunofluorescence (IF) and RNA fluorescence in situ hybridization (RNA FISH; how HCR™ IF + HCR™ RNA-FISH works)

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HCR™ IF + HCR™ RNA-FISH: Protocol Overview

Protein Detection Stage

RNA Detection Stage

Amplification Stage

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The same 3-stage protocol is used independent of the number of protein and RNA targets.

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Quantitative 1-step enzyme-free HCR™ signal amplification for all protein and RNA targets simultaneously

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Simple Robust Protocols

HCR™ IF + HCR™ RNA-FISH protocols are simple, robust, and enzyme-free, requiring only 3 stages independent of the number of protein and RNA targets.

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Straightforward Multiplexing

HCR™ enables straightforward multiplexing using 1-step quantitative signal amplification for all protein and RNA targets simultaneously.

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Any organism across the tree of life

Free custom probe design for any target mRNA in any organism. For protein targets, plug-and-play using your own 1º antibodies of choice (without modification) in combination with validated HCR™ 2º antibody probes.

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Normal Sleep Schedule

We favor protocols with an overnight probe incubation step to enable researchers to maintain a normal sleep schedule; durations can be customized as desired based on sample thickness.

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Simultaneous Quantitative High-Resolution Protein and RNA Imaging

Subcellular protein and RNA imaging enables protein and RNA relative quantitation with subcellular resolution in the anatomical context of highly autofluorescent samples.

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