Matchmaker Mammalian Assay Kit 2
Matchmaker Mammalian Assay Kit 2
Brand: Takara Bio.
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Matchmaker Mammalian Assay Kit 2
Complete kit for confirming a known protein-protein interaction using two-hybrid technology in mammalian cell culture. Protein-protein interaction is assayed by measuring SEAP gene expression. No lysis is required.
Overview
- Fast, convenient analysis of protein-protein interactions in mammalian cells
- SEAP secreted reporter eliminates the need for cell lysis
- Biologically relevant results are more likely in a mammalian system
Applications
- Confirm protein-protein interactions in mammalian cells
- Map interacting domains
The mammalian two-hybrid principle
The mammalian two-hybrid principle. The bait protein is fused to the DNA binding domain from GAL4 and the prey protein is fused the transcriptional activation domain of VP16. If the two proteins interact at the PGAL4-E1b promoter (GAL promoter), SEAP is secreted into the growth medium.
Chemiluminescent detection of protein-protein interactions
Chemiluminescent detection of protein-protein interactions. The bait vector pM-53 expresses p53 fused to the GAL4 DNA-binding domain. When HEK 293 cells were cotransfected with this bait together with the SV40 large T antigen prey vector, pVP16-T, strong expression of SEAP was detected because large T antigen interacts with p53. The CP protein, which does not interact strongly with p53, was used as a negative control.
The Matchmaker Two-Hybrid Assay Kit 2 procedure
The Matchmaker Two-Hybrid Assay Kit 2 procedure.
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The mammalian two-hybrid principle
The mammalian two-hybrid principle. The bait protein is fused to the DNA binding domain from GAL4 and the prey protein is fused the transcriptional activation domain of VP16. If the two proteins interact at the PGAL4-E1b promoter (GAL promoter), SEAP is secreted into the growth medium.
Chemiluminescent detection of protein-protein interactions
Chemiluminescent detection of protein-protein interactions. The bait vector pM-53 expresses p53 fused to the GAL4 DNA-binding domain. When HEK 293 cells were cotransfected with this bait together with the SV40 large T antigen prey vector, pVP16-T, strong expression of SEAP was detected because large T antigen interacts with p53. The CP protein, which does not interact strongly with p53, was used as a negative control.
The Matchmaker Two-Hybrid Assay Kit 2 procedure
The Matchmaker Two-Hybrid Assay Kit 2 procedure.
