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.
Cross-reactivity between the antibodies and any of the other analytes in this panel is non-detectable or negligible.
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)
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MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany
xMAP is a registered trademark of Luminex Corp
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