Protein phosphorylation represents a key 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 changes in both phosphorylation events, as well as changes due to total protein levels. 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 p38 kit has been developed for the simultaneous detection of phosphorylated p38 (Thr180/Tyr182) and total p38 in a single well using the Luminex system. According to the original vendor′s antibody information sheets, phospho-p38 antibody pair detects p38α, p38β and p38γ but not p38-δ, while the total p38 antibody pair detects only p38α. The detection assay is a rapid, convenient alternative to Western Blotting and immunoprecipitation procedures for the analysis of cell lysate samples. Each kit has sufficient reagents for one 96 well plate assay.The MILLIPLEX® 2-Plex Phospho/Total p38 magnetic bead kit has been developed for the simultaneous detection of phosphorylated p38 (Thr180/Thr182) and total p38 in a single well using the Luminex® system. This detection assay is a rapid, convenient alternative to Western Blotting and immunoprecipitation procedures for the analysis of cell lysate samples.
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 or total pathway proteins in tissue and cell lysate samples.An overnight (4°C) incubation is recommended for best results.This assay requires 25 μL diluted, filtered cell lysate per well.This kit must be run using the provided Assay Buffer.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:p38 (Total);p38 (Thr180/Tyr182)
Storage and Stability
Recommended storage for kit components is 2 - 8°C.
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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