ATF2 ELISA Kit

Catalog No. ABIN2866259

Human ATF2 ELISA Kit Colorimetric assay for quantification of Human ATF2.

Quick Overview for ATF2 ELISA Kit (ABIN2866259)

Target: ATF2 (Activating Transcription Factor 2 (ATF2))

Reactivity: Human

Detection Method: Colorimetric

Method Type: DNA-Binding ELISA

Application: ELISA

Sample Type: Cell Extracts, Tissue Samples

Product Details ATF2 ELISA Kit

Purpose: DNA-binding ELISA that facilitate the study of transcription factor activation in mammalien tissue and cell culture extracts.

Brand: TransAM®

Analytical Method: Quantitative

Specificity: TransAM ATF-2 Kits are tested for sensitivity in detecting ATF-2 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 untreated and treated HeLa (Anisomycin) cells are diluted to 0.625 μg/well and assayed using the TransAM ATF-2 Kit. The ratio of the signals from the treated cells over the untreated must be above 4. Lot No. 25209003 was developed for 7 minutes. It gave a ratio of 4.1 (Figure 1). The endogenous level of ATF-2 expression, and this ratio may vary depending on the cell type tested and the treatment used. TransAM ATF-2 Kits are also tested for specificity in detecting ATF-2 activity. TransAM ATF-2 assays are performed in the presence of an excess of oligonucleotide containing a wild-type or mutated ATF-2 consensus binding site (Figure 2). At 20X excess, the wild-type oligonucleotide prevents ATF-2 binding to the probe immobilized on the plate. Conversely, the mutated oligonucleotide has no effect on ATF-2 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 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 ATF2

Target: ATF2 (Activating Transcription Factor 2 (ATF2))

Alternative Name: Atf-2

Pathways: MAPK Signaling, RTK Signaling, Thyroid Hormone Synthesis, Activation of Innate immune Response, Chromatin Binding, Myometrial Relaxation and Contraction, Synaptic Membrane, Tube Formation, Toll-Like Receptors Cascades