What question did this study set out to answer?

To determine the cause of fluorescent reporter protein signal loss during nuclear protein staining and establish an optimized protocol to preserve cytosolic fluorescence.

February 19, 2014Cytometry Part A

Improved method to retain cytosolic reporter protein fluorescence while staining for nuclear proteins

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To determine the cause of fluorescent reporter protein signal loss during nuclear protein staining and establish an optimized protocol to preserve cytosolic fluorescence.
Evaluated standard transcription factor staining and fixation workflows to identify mechanisms underlying fluorescent protein signal loss.
Developed and optimized a fixation and permeabilization procedure to prevent reporter protein leakage while enabling antibody access to intracellular targets.
Demonstrated that standard nuclear staining protocols do not chemically destroy fluorescent proteins, but rather cause them to leak out due to insufficient fixation prior to permeabilization.
Established a reliable protocol that maintains cytosolic fluorescent protein retention while simultaneously enabling efficient staining of transcription factors and cytokines.

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Implication

Methodological study demonstrates enhanced fluorescent reporter retention during nuclear staining in permeabilized cells, indicating improved detection of transcription factors.

Cite This Study

Heinen et al. (2014) studied this question.

synapsesocial.com/papers/6a71611bfebe604dd70a0b80 https://doi.org/10.1002/cyto.a.22451

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Implication

Methodological study demonstrates enhanced fluorescent reporter retention during nuclear staining in permeabilized cells, indicating improved detection of transcription factors.

Improved method to retain cytosolic reporter protein fluorescence while staining for nuclear proteins

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Discussion

Implication

Cite This Study

Improved method to retain cytosolic reporter protein fluorescence while staining for nuclear proteins

Key Points

Discussion

Implication

Cite This Study

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