 |
NF-kB/p50 [3H] DNA binding assay
More about:
Assay concept>
1536-well assay protocol>
Reagent usage in 96-, 384- well formats>
Typical data>
Ordering information>
Assay concept
Return to top
1536-well assay protocol
- 15 µl binding buffer containing: 10 mM Hepes, pH 7, 20 mM sodium acetate, 0.2 mM EDTA, 0.05% (v/v) Nonidet P40
- 5 µl 50 mM DTT
- 5 µl 10 mg/ml BSA
- 5 µl 780 µCi/mmole [3H]DNA
- 5 µl 10 nM p50 protein
- Incubate at room temperature for 30 minutes
- 5 µl (0.5 mg) anti-GST antibody (27-4577-01)
- 5 µl (0.5 mg) polystyrene protein A bead (RPNQ0264)
- Incubate at room temperature for 60 minutes
- Image for 5 minutes
Return to top
Reagent usage in 96- and 384-well format
Assay components | 96-well | 384-well |
Assay volume | 100 µl | 50 µl |
Bead | 1 µg | 0.5 µg |
Antibody amount | 1.5 µg | 0.5 µg |
p50/p65 protein | 10 nM | 10 nM |
Protein amount | 1 pmole | 0.5 pmoles |
[3H]DNA | 0.4 pmoles | 0.2 pmoles |
|
Return to top
Typical Data
Return to top
Ordering information
The following products were used in this assay and can be purchased online
Return to top
Learn more about the LEADseeker Imaging System, begin with an introduction
Find out how to interpret your complex, screening data - visit the software and informatics pages.
View the entire range of reagents for the LEADseeker Imaging Systems
|
|
