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 2 (provided).1 - 25 µg cell lysate/well (recommended starting concentration is 40 to 1,000 µg protein/mL).Analytes available:eIF-4G (Ser1108);elF-4E (Ser209);elF-4B (Ser422);elF-2a (Ser51);4E-BP1 (Total);4E-BP1 (Thr37/46)

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

Protein synthesis is required for cellular growth and development. Normal cells grow in a controlled manner in response to environmental and developmental cues. However, cancer cells can reprogram cellular metabolism, favoring uncontrolled growth and survival. This can be achieved by altering signaling pathways that control cellular processes such as protein synthesis.The phosphorylation state of proteins involved in translation initiation is a limiting factor that regulates the formation or activity of translational complexes. In cancer cells, hyper-activated signaling pathways influence translation, allowing uncontrolled growth and survival. In addition, several components of translation initiation have been found to be mutated, post-translationally modified, or differentially expressed, and thus have been shown to act as oncogenes.Translational alterations can increase the overall rate of protein synthesis as well as activate regulatory mechanisms leading to the translation of specific messenger RNAs for proteins that promote cancer progression and survival.Each MILLIPLEX^® cell signaling kit includes: Stimulated and unstimulated cell lysates provided to qualify assay performance Premixed magnetic beads to capture analytes of interest Optimized detection antibody cocktails designed to yield consistent analyte profiles within a panelThe MILLIPLEX^® Protein Translation 6-plex Magnetic Bead kit is used to detect changes in phosphorylated eIF2a (Ser51), eIF-4B (Ser422), eIF-4E (Ser209), eIF-4G (Ser1108) and 4E-BP1 (Thr37/46), as well as total protein levels of 4E-BP1 in cell lysates using the Luminex^® system. The detection assay is a rapid, convenient alternative to Western Blotting and immunoprecipitation procedures. Each kit has sufficient reagents for one 96-well plate assay.

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.