Ordering Information
- : 70-601
- : 192 assays
Product Images
Relative light unit values obtained using Upstate's EZ-TFA Universal Transcription Factor Assay Kit may be compared with known standards or other test...
Universal EZ-TFA Chemiluminescent Transcription Factor Assay
Description:
Universal EZ-TFA Chemiluminescent Transcription Factor Assay
Trade Name:
Upstate (Millipore)
Qty/Pk:
192 assays
Background Information:
The UPSTATE Non-Radioactive Universal EZ-TFA Transcription Factor Assay provides a fast, sensitive method to detect specific transcription factor DNA binding activity in whole cell or nuclear extracts. This assay combines the principle of the electrophoretic mobility shift assay (EMSA) with the 96-well based enzyme-linked immunosorbent assay (ELISA), enabling manual or high-throughput sample analysis with greater sensitivity than conventional EMSA assays. After certain parameters are optimized, the entire assay takes less than 4 hours to complete with minimal hands-on time. The versatile set up allows for a flexible assay design and because the binding reaction occurs in solution, various parameters can be optimized such as probe titration, competitive oligonucleotide concentration, or treatment conditions. This kit contains the reagents and instructions necessary to develop an assay to monitor the DNA binding activity of any transcription factor of interest, or as a kit component for Upstate developed target specific Transcription Factor Assays. The Universal kit also allows for the design of native promoter capture probe sequences to interrogate specific transcription factor binding.
Test Principle:
The Non-Radioactive Universal Transcription Factor Assay kit is provided in a 96-well format. During the assay, the capture probe, a double stranded biotinylated oligonucleotide containing the consensus sequence for the specific transcription factor under study, is mixed with cellular (nuclear) extract in the transcription factor assay buffer provided directly in the streptavidin coated plate. When incubated together, the transcription factor contained in the extract binds to its consensus sequence. The biotinylated double stranded oligonucleotide bound to the transcription factor being studied is immobilized on the streptavidin-coated plate and any inactive, unbound material is washed away. The bound transcription factor is detected with a specific primary antibody. An HRP-conjugated secondary antibody is then used for detection and provides sensitive chemiluminescent detection that can be read in a microplate luminometer or by a CCD camera-coupled imaging system.
Test Principle:
The Non-Radioactive Universal Transcription Factor Assay kit is provided in a 96-well format. During the assay, the capture probe, a double stranded biotinylated oligonucleotide containing the consensus sequence for the specific transcription factor under study, is mixed with cellular (nuclear) extract in the transcription factor assay buffer provided directly in the streptavidin coated plate. When incubated together, the transcription factor contained in the extract binds to its consensus sequence. The biotinylated double stranded oligonucleotide bound to the transcription factor being studied is immobilized on the streptavidin-coated plate and any inactive, unbound material is washed away. The bound transcription factor is detected with a specific primary antibody. An HRP-conjugated secondary antibody is then used for detection and provides sensitive chemiluminescent detection that can be read in a microplate luminometer or by a CCD camera-coupled imaging system.
Key Applications:
- ELISA
- Activity Assay
Species Reactivity:
All
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Quality Assurance:
Routinely evaluated with gene specific target componet with all necessary controls.
Components:
- Streptavidin Coated Chemiluminescent Plate, Catalog # 90494.
- Blocking Reagent, Catalog # 90273.
- 10X Annealing Buffer, Catalog # 2007044
- Chemiluminescent Detection Reagent, Catalog #90495
- Chemiluminescent Reaction Buffer, Catalog #90496
- TFA Buffer, 5X, Catalog # 90272
- Gt X Rb, HRP Secondary Antibody, Catalog # 90276
- Gt X Ms, HRP Secondary Antibody, Catalog # 2003482
Brand Family:
Upstate
Storage Conditions:
Upon receipt, store components at the temperatures indicated on the labels.
Store at 4°C:
Transcription Factor Assay Chemiluminescent Plate, Catalog # 90494.
Blocking Reagent, Catalog # 90273.
10X Annealing Buffer, Catalog # 2007044.
Chemiluminescent Detection Reagent, Catalog #90495.
Chemiluminescent Reaction Buffer, Catalog #90496.
Store at -20°C:
Gt X Rb, HRP Secondary Antibody, Catalog # 90276.
TFA Buffer, 5X, Catalog # 90272
Gt X Ms, HRP Secondary Antibody, Catalog # 2003482.
Store at 4°C:
Transcription Factor Assay Chemiluminescent Plate, Catalog # 90494.
Blocking Reagent, Catalog # 90273.
10X Annealing Buffer, Catalog # 2007044.
Chemiluminescent Detection Reagent, Catalog #90495.
Chemiluminescent Reaction Buffer, Catalog #90496.
Store at -20°C:
Gt X Rb, HRP Secondary Antibody, Catalog # 90276.
TFA Buffer, 5X, Catalog # 90272
Gt X Ms, HRP Secondary Antibody, Catalog # 2003482.
Detection Methods:
Chemiluminescence
Product Name:
Universal EZ-TFA Chemiluminescent Transcription Factor Assay
Materials Required but Not Delivered:
Reagents Cells, stimulated or treated as needed for the experimental system Capture probe Specific competitor Primary antibody Nuclear extraction kit (Catalog No. 2900) Nuclease free water (Catalog No. 3098)