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Native purification of transcription factors with μMACS™ Technology.
Functional isolation of any DNA-binding protein by using a specific DNA target sequence.
90-minute protocol with ready-to-use buffers but without ultracentrifugation.
Detection of novel transcription factors and analysis of target sequences. The μMACS FactorFinder Kit enables the highly specific magnetic isolation of DNA-binding proteins. The applications of the kit are as versatile as the questions in transcriptional regulation: By protein isolation individual transcription factors and their cofactors can be detected and characterized. Thereby, complete profiles are generated. In addition, DNA-protein interactions and target sequences can be analyzed. Furthermore, the kit facilitates the investigation of transcription factors as potential therapeutic targets.
Fast and functional. In about 45 minutes, a whole cell lysate containing both the cytosolic and the nuclear fraction is prepared with the provided buffers—but no tedious ultracentrifugation steps. Within another 45 minutes, the native transcription factor of interest is magnetically isolated.
MACS® Technology for isolation of transcription factors. The addition of biotinylated target DNA and magnetizable μMACS Streptavidin MicroBeads to the lysate magnetically labels the target protein. The mixture is loaded into the μ Column which is placed in the magnetic field of a MACS® Separator. Application of wash buffers removes unspecific molecules. The final elution step yields native transcription factor that is suitable for functional downstream assays. Alternatively, a denaturing elution provides protein ready for SDS-PAGE analysis.
In contrast to conventional gel shift assays and ELISA-based methods, the innovative μMACS FactorFinder Kit offers a system not only for detection but also for isolation of transcription factors; thereby, facilitating signal transduction studies in for example immunology, oncology, or stem cell research. |
| Columns |
| μ Columns (included in kit) |
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| Figure 1 |
| Working scheme for the isolation of transcription factors. |
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| Figure 2 |
Isolation of phosphorylated STAT-3 (pSTAT-3) from activated T cells.
T cells from human PBMCs were activated and expanded in the presence of IL-2 using the T Cell Activation/Expansion Kit. After three days, cells were incubated with IL-15 for 15 min, washed and lysed with Cell Lysis Buffer. Whole cell lysate was incubated with a biotinylated 31 bp DNA probe comprising the STAT-3 binding sequence, μMACS Streptavidin MicroBeads were added, and the magnetic isolation was performed. The transcription factor was eluted with Native Elution Buffer. The Western blot (using a phosphospecific STAT-3 antibody) shows a high amount of pSTAT-3 in the eluate. L: cell lysate; FT: flow-through; W1, W2: wash fractions; E: eluate. |
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| Products |
| μMACS FactorFinder Kit |
- for 20 isolations
Components
- 2 mL μMACS Streptavidin MicroBeads
- 5 mL Cell Lysis Buffer
- 12 mL Binding Buffer
- 10 mL Wash Buffer LS
- 10 mL Wash Buffer HS
- 4 mL Native Elution Buffer
- 0.5 mL Binding Enhancer (100× stock solution)
- 20 μ Columns
Download data sheet
130-092-317
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| μMACS FactorFinder Starting Kit |
- for 20 isolations
Components
- 1 μMACS FactorFinder Kit
- 1 μMACS Separator
- 1 MACS MultiStand
Download data sheet
130-092-318
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