STATs (Signal Transducers and Activators of Transcription) are used by receptors for a wide variety of ligands including cytokines, hormones, growth factors and neurotransmitters. The phosphorylation of STAT proteins at conserved tyrosine residues activates SH2-mediated dimerization followed rapidly by nuclear translocation. STAT dimers bind to IRE (interferon response element) and GAS (gamma interferon-activated sequence) DNA elements, resulting in the transcriptional regulation of downstream genes. STAT pathways play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses and stem cell differentiation.Protein phosphorylation represents the major mechanism used in regulating cellular functions in all eukaryotic cells. Aberrant phosphorylation has been implicated in the onset and development of many diseases including metabolic disorders, inflammatory disease, cancer, etc. Changes in protein phosphorylation can be attributed to both changes in phosphorylation events as well as changes due to total protein levels. In order to distinguish the changes in phosphorylation from changes in protein expression, it is important to normalize the signal from phosphorylation over the signal from total protein. For this need, the MILLIPLEX® MAP 2-plex Phospho/Total STAT3 Kit (Cat. No. 48-623MAG) has been developed for the simultaneous detection of phosphorylated STAT3 (Tyr705) and total STAT3 in a single well using the Luminex system.
Cross-reactivity between the antibodies and any of the other analytes in this kit is non-detectable or negligible.
Intracellular Bead-Based Multiplex Assays using the Luminex technology enables the simultaneous relative quantitation of multiple phosphorylation or total pathway proteins in tissue and cell lysate samples. The analytes available for this multiplex kit are: STAT3 (Total), STAT3 (Tyr705).This is an overnight (4°C) incubation assay.This kit must be run using the provided Assay Buffer.This assay requires 25 μL diluted filtered cell lysate per well.The suggested working range of protein concentration for the assay is 1 to 25 μg of total protein/well (25 μL/well at 40 to 1,000 μg/mL).Analytes available:STAT3 (Total)STAT3 (Tyr705)
Luminex is a registered trademark of Luminex Corp
MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany
xMAP is a registered trademark of Luminex Corp
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