TransAM® STAT3

by Active Motif
Available SKUs: 45696 |
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Description

TransAM® Kits are DNA-binding ELISAs that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. Assays are available for over 40 different targets (see the list at right). 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. For complete details, click the TransAM® Method tab below.

TransAM® STAT Transcription Factor ELISA Kits

TransAM STAT Kits provide everything needed to study Signal Transducers and Activators of Transcription (STAT), including a positive control extract. TransAM STAT3 Kits are available with antibodies specific for the activated form of STAT3. A TransAM STAT Family kit is also available with the ability to independently screen the members of the STAT family (STAT1?, STAT3, STAT5A & STAT5B) simultaneously. The STAT1?, 5A and 5B antibodies can be used with human and rat extracts, while the STAT3 antibodies can be used with human, mouse and rat extracts. See the STAT Info tab below for kit data and more information; the kit manual can be downloaded under the Documents tab.

 
Name Format Cat No. Price  
TransAM® STAT Family 2 x 96 rxns 42296 $1,145 Buy Now
TransAM® STAT3 1 x 96 rxns 45196 $685 Buy Now
TransAM® STAT3 5 x 96 rxns 45696 $2,795 Buy Now

STAT Transcription Factor Info

Signal Transducers and Activators of Transcription (STAT) are a family of latent cytoplasmic proteins that are activated in response to cytokines and growth factors. The STAT family has seven members: STAT1, 2, 3, 4, 5A, 5B and 6. STATs are involved in many pathways that regulate cellular functions in the lympho-hematopoietic system. STATs can be activated by ligands such as interferons (IFNs), epidermal growth factor (EGF), platelet derived growth factor (PDGF) and interleukins (IL). When members of the STAT family are phosphorylated they form homo- or heterodimers and translocate to the nucleus to activate transcription. STAT1? homodimerizes upon tyrosine or serine phosphorylation induced by IFN?, but forms heterodimers with STAT2 when induced by IFN?/?. STAT3 is activated by phopshorylation of tyrosine 705 and serine 727 to mediate the expression of genes involved in cell growth and apoptosis. STAT5A and STAT5B are activated in response to several cell ligands and growth hormones. They are capable of serving as both transcriptional activators and signal transducers.

 

 
 
Figure 1: Monitoring STAT Family member activation using the TransAM STAT Family Kit.

STAT1?, STAT3, STAT5A and STAT5B activation were assayed using the TransAM STAT Family Kit. 1:1000 dilutions of each antibody in the kit were tested using 5 µg/well of nuclear extract prepared from a stimulated cell line: STAT1? was tested with U-937 (TPA + IFN?), STAT3 with Hep G2 (IL-6, 100 ng/ml), and STAT5A and STAT5B with Nb2 (prolactin). Assays were performed in the absence or presence of 20 pmol of competitor oligonucleotide that contains either a wild-type or mutated STAT consensus binding site. Note that the wild-type oligonucleotide reduces STAT binding by over 90%, while incubation with the mutant STAT competitor oligo has a limited effect on STAT 1?, STAT3, STAT5A or STAT5B binding to DNA.

The TransAM® transcription factor ELISA advantage

Historically, transcription factor studies have been conducted using gelshift, Western blot and reporter plasmid transfections, which are time-consuming, do not allow for high-throughput and provide only semi-quantitative results. TransAM assays are up to 100 times more sensitive than gelshift techniques, and can be completed in less than 5 hours. Because T

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