C/ebp A/b ELISA Kit
Reactivity: Human
Colorimetric
DNA-Binding ELISA
Cell Extracts, Tissue Samples
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- Target
- C/ebp A/b
- Reactivity
- Human
- Detection Method
- Colorimetric
- Method Type
- DNA-Binding ELISA
- Application
- ELISA
- Purpose
- DNA-binding ELISA that facilitate the study of transcription factor activation in mammalien tissue and cell culture extracts.
- Brand
- TransAM®
- Sample Type
- Cell Extracts, Tissue Samples
- Analytical Method
- Quantitative
- Specificity
- TransAM C/EBP a/b Kits are tested for sensitivity in detecting both C/EBPa and C/EBPb using nuclear extracts from Jurkat cells and U-937 cells.
- Characteristics
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Transcription factors are DNA-binding proteins that tightly regulate gene expression. They consist of two distinct domains - one that displays high affinity for a specific DNA sequence and one that confers transcriptional activity. Transcription factors are activated by phosphorylation of specific residues or by processing bound inhibitory proteins. Understanding and quantifying transcription factors is essential for the study of cell functions in relation to differentiation, brain activity, immune response, inflammation and various disease states. TransAM® Kits are sensitive, non-radioactive transcription factor ELISA kits that facilitate the study of transcription factor activation in mammalian tissue and cell extracts.
TransAM® Kits are DNA-binding ELISAs that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. Each kit includes a 96-stripwell plate in which multiple copies of a specific double-stranded oligonucleotide have been immobilized. When nuclear or whole-cell extract is added, activated transcription factor of interest binds the oligonucleotide at its consensus binding site and is quantified using the included antibody, which is specific for the bound, active form of the transcription factor being studied. - Components
- One or five 96-well plate(s) with plate sealer(s), primary antibody, HRP-conjugated secondary antibody, wild-type and mutated competitor oligonucleotides, positive control cell extract, DTT, Protease Inhibitor Cocktail, Lysis, Binding, 10X Washing and 10X Antibody Binding Buffers, and Developing and Stop Solutions.
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- Application Notes
- Optimal working dilution should be determined by the investigator.
- Comment
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These extracts are diluted down to 0.625 μg/well and assayed using the TransAM C/EBP a/b Kit. The ratio of the signals from the U-937 over the Jurkat cell extracts must be above 4 at 10 μg/well. For Lot No.34408005, C/EBP alpha was developed for 10 minutes with a ratio of 5, while C/EBP beta was developed for 3 minutes with a ratio of 5 (Figures 1a & 1b). TransAM Kits are also tested for specificity. Assays are performed with an excess of oligonucleotide containing a wild-type or mutated C/EBP consensus binding site. At 40 pmol, the wild-type oligo prevents C/EBP binding to the probe immobilized on the plate, while the mutated oligonucleotide has little effect on binding (Figure 2, data shown only for C/EBPb).
- Assay Time
- 5 h
- Plate
- Pre-coated
- Restrictions
- For Research Use only
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- Storage
- 4 °C/-20 °C/-80 °C
- Storage Comment
- Except for the cell extract that must be kept at -80°C, the kit components can be stored at -20°C prior to first use. Then, we recommend storing the kit at 4°C except for the oligonucleotides, DTT, Herring sperm DNA and Protease Inhibitor Cocktail that should be kept at -20°C, and the cell extract at -80°C. This product is guaranteed for 6 months from date of receipt.
- Expiry Date
- 6 months
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- Target
- C/ebp A/b
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