T-Bet ELISA Kit

Catalog No. ABIN2866295

Product Details T-Bet ELISA Kit

Target: T-Bet

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 T-bet Kits are tested for sensitivity in detecting T-bet activation.

Characteristics: 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.

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

Nuclear extracts prepared from Cos-7 cells that are either transfected with T-bet or not and stimulated with PMA/ionomycin. Nuclear extracts are diluted to 0.156 μg/well and assayed using the TransAM T-bet Kit. The ratio of the signals from the stimulated over unstimulated cells must be above 4. Lot No. 29408001 was developed for 3 minutes. It gave a ratio of 120 (Figure 1). This ratio may vary depending on the level of T-bet activation in a given cell type. TransAM T-bet Kits are also tested for specificity in detecting T-bet activation. TransAM T-bet assays are performed in the presence of an excess of oligonucleotide containing a wild-type or mutated T-bet consensus binding site (Figure 2). At 40X excess, the wild-type oligonucleotide prevents T-bet binding to the probe immobilized on the plate. Conversely, the mutated oligonucleotide has no effect on T-bet binding.

Assay Time: 5 h

Plate: Pre-coated

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 °C/-80 °C

Storage Comment: Store the cell extract at -80°C. Other 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, poly [d(I-C)] and Protease Inhibitor Cocktail that should be kept at -20°C. This product is guaranteed for 6 months from date of receipt.

Expiry Date: 6 months

Target Details for T-Bet

Target: T-Bet

Abstract: T-Bet Products

Synonyms: T-PET ELISA Kit, T-bet ELISA Kit, TBET ELISA Kit, TBLYM ELISA Kit, TBT1 ELISA Kit, Tbet ELISA Kit, Tblym ELISA Kit, T-box 21 ELISA Kit, TBX21 ELISA Kit, Tbx21 ELISA Kit