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- Specific transcription factor-DNA binding assay
- Perfect alternative to EMSA
- Easy to perform in an ELISA format
- Non-radioactive assay
- High throughput (96-well plate format)
- Assay can be completed within 5 hours
Target Protein Information
- 96-well Strip Microplate pre-coated with DNA probes
- DNA Binding Buffer
- Positive Control Sample
- Specific Competitor DNA probe
- Non-specific Competitor DNA probe
- Assay Reagent
- Wash Buffer
- Primary Antibody
- HRP-conjugated Secondary Antibody
- TMB One-Step Substrate Reagent
- Stop Solution
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Absorbent paper
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Benchtop rocker or shaker
- Microplate reader capable of measuring absorbance at 450 nm
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared HRP-secondary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-a. After stimulation with TNF-a, activated NF-κB p65 is translocated into the nucleus where it binds with its corresponding DNA.
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-a with the specific competitor or non-specific competitor. The result shows specific binding of NF-κB p65 to the NF-κB DNA binding site.
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells treated with TNF-a and purified NF-κB p50. No cross-reactivity has been observed between NF-κB p65 and p50.
Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.
- 1, 2, or 5 x 96-Well Strip Microplate Kit
- IGF Signaling Pathway
- NFkB Signaling Pathway
- PI3K Signaling Pathway
- PKC-Theta Signaling Pathway
- B Cell Receptor
- JAK/STAT Signaling Pathway
- MAPK Signaling Pathway
- mTOR Signaling Pathway
- T Cell Receptor
- 96-well Microplate
- Nuclear Extracts
- Cell Lysates
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.