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STATs (signal transducers and activators of transcription) are a family of latent cytoplasmic proteins that are activated when cells encounter various extracellular polypeptides, such as interferons and interleukines. STATs are involved in many pathways that regulate cellular functions in the lympho-hematopoietic system. The STAT family has seven members: STAT1, 2, 3, 4, 5A, 5B and 6. STAT1 forms heterodimers with STAT2 and 3; STAT4 and 5 only form homodimers. Tyrosine phosphorylation around residue 700 is essential for the dimerization of STATs and the concomitant nuclear translocation of the dimer. STAT dimers and heterodimers, but not monomers, are competent to bind DNA.
TransAM STAT Kits are available in two different formats. TransAM STAT3 contains antibody specific for activated STAT3. The TransAM STAT Family Kit contains antibodies specific for the active forms of STAT1α, STAT3, STAT5A and STAT5B when they are bound to the target DNA.
Applications
TransAM STAT Kits can be used in investigations of the pathophysiology of many prevalent diseases including heart attack, cancer, diabetes, asthma and other chronic lung diseases. Applications include:
- Drug screening and/or potency toward STAT activation
- JAK kinase studies
- Cytokine stimulation
- Protein structure/function studies
Figure 1: Monitoring STAT Family member activation using the TransAM STAT Family Kit.
STAT1α, 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: STAT1α was tested with U-937 (TPA + IFNγ), 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 1α, STAT3, STAT5A or STAT5B binding to DNA.
