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phospho-p38α (Thr180/Tyr182) STAR ELISA Kit
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PRODUCT FAMILY INFORMATION
STAR ELISA Kits
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
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MAPK Antibodies, Proteins, Inhibitors, & Assays
Offering hundreds of premium quality antibodies, enzymes, activity assays, and small molecule inhibitors for MAPK signaling research
EMD Millipore offers hundreds of premium quality antibodies, enzymes, activity assays, and small molecule inhibitors for MAPK signaling research. All EMD Millipore products are based on the expertise of Upstate, Chemicon, and Calbiochem.
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Description:
phospho-p38α (Thr180/Tyr182) STAR ELISA Kit
Trade Name:
Upstate (Millipore)
Qty/Pk:
96 assays
Background Information:
I. TEST PRINCIPLE
The UPSTATE® colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The p38 MAPK plate is coated with a specific mouse monoclonal p38 MAPK capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standard included in the kit are incubated in the microwells allowing p38 MAPK antigen to be captured in the plate wells. The plate is then washed to remove any non-bound unspecific material. The wells are then incubated with a specific rabbit anti-phospho-p38 (Thr180/Tyr182) antibody to detect the captured p38 (Thr180/Tyr182) on the plate well. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. This allows for a sensitive enzymatic detection of the sample. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader.
The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to develop a standard line of detection.
II. p38 BACKGROUND
The p38 MAPK family of Ser/Thr kinase proteins is made up of 4 isoforms, p38alpha (MAPK14 and SAPK2a), p38beta (MAPK11 and SAPK2b), p38delta (MAPK13, SAPK4), and p38gamma (MAPK12, SAPK3). The other p38 isoforms share 74%, 57%, and 60% sequence identity with the alpha isoform, respectively. All four isoforms posses the TGY motif and are activated by the dual phosphorylation of the threonine and tyrosine residues in this motif, which correspond to Thr180/Tyr182 in human p38alpha. p38, especially the alpha and beta isoforms, are able to phosphorylate a wide array of targets including numerous kinases including MAPKAP-K2, MAPKAP-K3, PRAK, MSK, and MNK as well as transcription factors including STAT1, p53, NFATp, ATF-2, Elk-1 (although weakly), and MEF2a/c. p38 MAPK family members have traditionally been associated with stress, immune response, and regulation of cell proliferation and apoptosis. They also have a role in regulating the production of pro-inflammatory cytokines and have been associated with diseases such as asthma and several autoimmune diseases, including rheumatoid arthritis, diabetes, and inflammatory bowel disease. Just as ERK is phosphorylated and activated by MEK, p38alpha MAP Kinase is activated predominantly by its upstream regulatory kinases MKK6 and MKK3 as well as MKK4 which also activates JNK, but is not a substrate for MKK7 like JNK i |