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Molecular Interaction Research

Answering "do these two molecules interact directly." Two parallel tracks — protein-protein and protein-DNA — both follow the same logic: point-to-point initial screening, then precise validation.

TECHNICAL ROUTE

Technical Route

Protein-Protein Interaction Protein-DNA Interaction 1 Yeast Two-Hybrid Targeted Screening of Candidate Interactions 2 Co-IP/Pull-down/BiFC Precise Validation of Interaction 1 Yeast One-Hybrid Targeted Screening of Candidate Interactions 2 EMSA/DNA Pull-down/ChIP Precise Validation of Binding 3 Dual-Luciferase Validation Confirming Effect on Transcriptional Activity Tip: Use high-throughput methods like yeast hybrid assays to narrow candidates first, then validate individually with Co-IP / EMSA.
RESEARCH STEPS

Research Steps & Corresponding Services

1

Protein-Protein Interaction: Initial Screening

Use point-to-point yeast two-hybrid validation to make an initial determination of whether two candidate proteins interact.

Yeast Two-Hybrid →
2

Protein-Protein Interaction: Precise Validation

Further and precisely validate the interaction in cell lysate or live cells with orthogonal methods such as Co-IP, Pull-down, or BiFC.

Co-IP (Co-Immunoprecipitation) →Pull-down Assay →BiFC (Bimolecular Fluorescence Complementation) →
3

Protein-DNA Interaction: Initial Screening

Use point-to-point yeast one-hybrid validation to make an initial determination of whether a candidate protein (e.g. a transcription factor) binds a specific DNA cis-element.

Yeast One-Hybrid →
4

Protein-DNA Interaction: Precise Validation

Use an EMSA gel-shift assay to precisely validate the in vitro binding relationship between the protein and DNA, and confirm binding specificity.

EMSA (Gel Shift Assay) →
5

Confirm Effect on Transcriptional Activity

Use a dual-luciferase reporter assay to confirm the actual regulatory effect of this interaction on downstream gene transcriptional activity, completing the validation loop from "binding" to "function."

Dual-Luciferase Reporter Assay →

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OTHER SOLUTIONS

Other Solutions

Gene Function Research
Molecular Mechanism Research
Molecular Interaction Research
Expression Regulation Research
Non-coding RNA Research
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