Key features and details
- Assay type: Cell-based (quantitative)
- Detection method: Luminescent
- Sample type: Adherent cells, Suspension cells
- Reacts with: Mouse, Rat, Human
Product namePhospho-p38 MAPK (T180/Y182) + Total In-Cell ELISA Kit (Chemiluminescent)
See all p38 kits
Sample typeAdherent cells, Suspension cells
Assay typeCell-based (quantitative)
Assay durationMultiple steps standard assay
Species reactivityReacts with: Mouse, Rat, Human
Phospho-p38 MAPK (T180/Y182) + Total In-Cell ELISA Kit (Chemiluminescent) (ab207483) provides a simple, efficient, cell-based method to monitor proteins activated by phosphorylation. The kit is designed specifically to quantify activated (phosphorylated) p38 MAPK and/or total p38 MAPK. Cells are cultured in 96-well plates and stimulated to induce the pathway of interest. Following stimulation, the cells are rapidly fixed to preserve activation-specific protein modifications. Each well is then incubated with a primary antibody that recognizes either phosphorylated p38 MAPK or total p38 MAPK. Subsequent incubation with secondary HRP-conjugated antibody and developing solution provides an easily quantified chemiluminescent readout. The relative number of cells in each well is then determined using the provided Crystal Violet solution. The 96-well plate format is suitable for high-throughput screening applications.
The Phospho-p38 MAPK (T180/Y182) + Total In-Cell ELISA Kit (Chemiluminescent) contains two 96-well plates and two primary antibodies. The phospho-p38 antibody was raised in rabbit against a synthetic phospho-peptide corresponding to residues flanking the phosphorylation sites Thr180/Tyr182 of human p38 MAP kinase. This antibody recognizes only phosphorylated p38 MAPK. The total-p38 antibody was raised in rabbit and recognizes p38 MAPK regardless of its phosphorylation state. The kit can be used to study phosphorylated p38 MAPK relative to cell number or to determine p38 MAPK phosphorylation relative to the total p38 MAPK protein found in the cells. Once the phospho-p38 MAPK and total-p38 MAPK signals have been normalized for cell number, a comparison of the ratio of phosphorylated p38 MAPK to total p38 MAPK for each of the cell growth conditions can be made. The provided total-p38 antibody can be used as a positive control to demonstrate that the cells contain p38 MAPK, the kit reagents are functional and that the protocol is performed correctly. Also, because fixed cells are stable for several weeks, many plates can be prepared simultaneously and then the assay performed when desired.
p38 MAPK is a member of the mitogen-activated protein kinase (MAPKs) family of serine/threonine protein kinases. These proteins are widely conserved among eukaryotes and are involved in many cellular processes such as cell proliferation, cell movement and cell death. MAPKs phosphorylate specific serines and threonines on target protein substrates and function in signaling cascades that convey external stimuli from the cell surface to cellular targets such as translational machinery, cytoskeletal proteins and transcription factors.
The p38 MAPK signaling cascade is initiated when external stimuli such as growth factors, cellular stress (e.g. osmotic shock, radiation) or inflammatory cytokines are recognized by cell-surface receptors. This results in the activation of the RHO family GTPases (Rac, Rho, Cdc42), which activate MAPKKKs (e.g. MLK, TAK, ASK1). These MAPKKKs in turn phosphorylate and activate p38 MAPK kinase, which then phosphorylates p38 MAPK. Targets of phosphorylated (activated) p38 MAPK include the transcription factors STAT1, Myc/Max, Elk-1, CHOP, MEF2, ATF-2 and (through Msk-1) CREB.
Aberrant regulation of p38 MAPK is thought to contribute to a variety of physiological pathologies, including human diseases such as asthma and autoimmunity.
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Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 5 x 96 tests 1% SDS Solution 1 x 22ml 5 x 22ml 10% Triton X-100 1 x 10ml 5 x 10ml 10X PBS 1 x 120ml 5 x 120ml 1X Antibody Blocking Buffer 1 x 22ml 5 x 22ml 1X Antibody Dilution Buffer 1 x 30ml 5 x 30ml 96-well tissue culture plate 2 units 10 units Anti-rabbit HRP-conjugated IgG 1 x 11µl 5 x 11µl Chemiluminescent Reagent 2 x 2ml 10 x 2ml Crystal Violet Solution 1 x 22ml 5 x 22ml Phospho-p38 antibody 1 x 9µl 5 x 9µl Plate sealer 2 units 10 units Reaction Buffer 2 x 4ml 10 x 4ml Total-p38 antibody 1 x 9µl 5 x 9µl
FunctionResponds to activation by environmental stress, pro-inflammatory cytokines and lipopolysaccharide (LPS) by phosphorylating a number of transcription factors, such as ELK1 and ATF2 and several downstream kinases, such as MAPKAPK2 and MAPKAPK5. Plays a critical role in the production of some cytokines, for example IL-6. May play a role in stabilization of EPO mRNA during hypoxic stress. Isoform Mxi2 activation is stimulated by mitogens and oxidative stress and only poorly phosphorylates ELK1 and ATF2. Isoform Exip may play a role in the early onset of apoptosis.
Tissue specificityBrain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.
Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain.
DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
modificationsDually phosphorylated on Thr-180 and Tyr-182, which activates the enzyme.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
- CSAID binding protein
- CSAID Binding Protein 1
- CSAID-binding protein
ab207483 has not yet been referenced specifically in any publications.