Application

Intracellular Bead-Based Multiplex Assays using the Luminex technology enables the simultaneous relative quantitation of multiple phosphorylation and total pathway proteins in tissue and cell lysate samples. Compare Multiplexing results to those of Western blotting.An overnight (4°C) incubation is recommended for best results.This assay requires 25 µL diluted cell lysate per well.This kit must be run using Assay Buffer 1 (provided).1 - 25 µg cell lysate/well (recommended starting concentration is 40 to 1,000 µg protein/mL).Analytes available:Akt (Ser473);ERK (Thr185/Tyr187);Smad2 (Ser465/Ser467);Smad3 (Ser423/Ser425);Smad4 (Total);TGFβRII (Total)

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Cells respond to their environment in many different ways through intracellular signaling. The TGFβ pathway plays a central role in a number of normal cellular processes including proliferation, differentiation, apoptosis, migration, adhesion, immune response and other functions in most cells. Although the TGFβ pathway regulates a wide range of processes, the pathway is fairly simple. TGFβ dimers bind to TGFβ Type II Receptor which recruits and phosphorylates TGFβ Type I Receptor. The Type I Receptor then recruits and phosphorylates SMAD2/3, which then binds to SMAD4 and forms a complex that enters the cell nucleus where it acts as a transcription factor for various genes. TGFβ Receptor activates the SMAD-dependent canonical pathway, as well as SMAD-independent non-canonical pathways, such as PI3K/Akt, ERK, JNK, and p38. Aberrant TGFβ signaling is involved in the development of multiple diseases, including hematological malignancies such as leukemia, and impaired wound healing, neurodegenerative diseases, developmental disorders and pulmonary hypertension. Loss-of-function mutations promote tumorigenesis by suppressing the immune system and the epithelial mesenchymal transition (EMT), although TGFβ signaling has also been implicated in tumor inhibition. Immune cells such as B cells, T cells, dendritic cells and macrophages secrete TGFβ, which downregulates their proliferation, differentiation and activation by other cytokines. Consequently, modifications in TGFβ signaling have been linked to autoimmune and inflammatory diseases and cancer.

Legal Information

xMAP is a registered trademark of Luminex Corp

Luminex is a registered trademark of Luminex Corp

MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

96-well plate

Specificity

Cross-reactivity between the antibodies and any of the other analytes in this panel is non-detectable or negligible.

Storage and Stability

Recommended storage for kit components is 2 - 8°C.