MAPK14 ELISA Kit (Rat) (OKCD02630)

Catalog No: OKCD02630

Size:96WELLS

Rat MAPK14 ELISA kit standard curve

Product Info

• Predicted Species Reactivity: Rat|Rattus norvegicus
• Application: Enzyme-linked Immunosorbent assay-Sandwich
• ELISA Kit Detection Method: Colorimetric
• ELISA Kit Duration: 3h
• ELISA Kit Principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mitogen Activated Protein Kinase 14 (MAPK14). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mitogen Activated Protein Kinase 14 (MAPK14). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mitogen Activated Protein Kinase 14 (MAPK14), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Mitogen Activated Protein Kinase 14 (MAPK14) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
• ELISA Kit Range: 0.156-10ng/mL
• ELISA Kit Reproducibility: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mitogen Activated Protein Kinase 14 (MAPK14) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mitogen Activated Protein Kinase 14 (MAPK14) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
• ELISA Kit Component:
  • Anti-MAPK14 Microplate: 96 Wells (12 x 8 Well strips)
  • MAPK14 Lyophilized Standard: 2
  • 100X Biotinylated MAPK14 Detector Antibody: 1 x 120 uL
  • 100X Avidin-HRP Conjugate: 1 x 120 uL
  • Standard Diluent: 1 x 20 mL
  • Detector Antibody Diluent: 1 x 12 mL
  • Conjugate Diluent: 1 x 12 mL
  • 30X Wash Buffer: 1 x 20 mL
  • Stop Solution: 1 x 6 mL
  • TMB Substrate: 1 x 9 mL
• Reconstitution and Storage: 2°C to 8°C|-20°C
• Sample Type: Tissue homogenates and other biological fluids
• Sensitivity: 0.43 ng/mL
• Specificity: This assay has high sensitivity and excellent specificity for detection of Mitogen Activated Protein Kinase 14 (MAPK14). No significant cross-reactivity or interference between Mitogen Activated Protein Kinase 14 (MAPK14) and analogues was observed
• Assay Info: Assay Methodology: Quantitative Sandwich ELISA

Target Info

• Gene Symbol: MAPK14
• Alias Symbols: CRK1;Mitogen-activated protein kinase p38 alpha.
• Protein Name: Mitogen-activated protein kinase 14
• Description of Target: Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK14 is one of the four p38 MAPKs which play an important role in the cascades of cellular responses evoked by extracellular stimuli such as proinflammatory cytokines or physical stress leading to direct activation of transcription factors. Accordingly, p38 MAPKs phosphorylate a broad range of proteins and it has been estimated that they may have approximately 200 to 300 substrates each. Some of the targets are downstream kinases which are activated through phosphorylation and further phosphorylate additional targets. RPS6KA5/MSK1 and RPS6KA4/MSK2 can directly phosphorylate and activate transcription factors such as CREB1, ATF1, the NF-kappa-B isoform RELA/NFKB3, STAT1 and STAT3, but can also phosphorylate histone H3 and the nucleosomal protein HMGN1. RPS6KA5/MSK1 and RPS6KA4/MSK2 play important roles in the rapid induction of immediate-early genes in response to stress or mitogenic stimuli, either by inducing chromatin remodeling or by recruiting the transcription machinery. On the other hand, two other kinase targets, MAPKAPK2/MK2 and MAPKAPK3/MK3, participate in the control of gene expression mostly at the post-transcriptional level, by phosphorylating ZFP36 (tristetraprolin) and ELAVL1, and by regulating EEF2K, which is important for the elongation of mRNA during translation. MKNK1/MNK1 and MKNK2/MNK2, two other kinases activated by p38 MAPKs, regulate protein synthesis by phosphorylating the initiation factor EIF4E2. MAPK14 interacts also with casein kinase II, leading to its activation through autophosphorylation and further phosphorylation of TP53/p53. In the cytoplasm, the p38 MAPK pathway is an important regulator of protein turnover. For example, CFLAR is an inhibitor of TNF-induced apoptosis whose proteasome-mediated degradation is regulated by p38 MAPK phosphorylation. In a similar way, MAPK14 phosphorylates the ubiquitin ligase SIAH2, regulating its activity towards EGLN3. MAPK14 may also inhibit the lysosomal degradation pathway of autophagy by interfering with the intracellular trafficking of the transmembrane protein ATG9. Another function of MAPK14 is to regulate the endocytosis of membrane receptors by different mechanisms that impinge on the small GTPase RAB5A. In addition, clathrin-mediated EGFR internalization induced by inflammatory cytokines and UV irradiation depends on MAPK14-mediated phosphorylation of EGFR itself as well as of RAB5A effectors. Ectodomain shedding of transmembrane proteins is regulated by p38 MAPKs as well. In response to inflammatory stimuli, p38 MAPKs phosphorylate the membrane-associated metalloprotease ADAM17. Such phosphorylation is required for ADAM17-mediated ectodomain shedding of TGF-alpha family ligands, which results in the activation of EGFR signaling and cell proliferation. Another p38 MAPK substrate is FGFR1. FGFR1 can be translocated from the extracellular space into the cytosol and nucleus of target cells, and regulates processes such as rRNA synthesis and cell growth. FGFR1 translocation requires p38 MAPK activation. In the nucleus, many transcription factors are phosphorylated and activated by p38 MAPKs in response to different stimuli. Classical examples include ATF1, ATF2, ATF6, ELK1, PTPRH, DDIT3, TP53/p53 and MEF2C and MEF2A. The p38 MAPKs are emerging as important modulators of gene expression by regulating chromatin modifiers and remodelers. The promoters of several genes involved in the inflammatory response, such as IL6, IL8 and IL12B, display a p38 MAPK-dependent enrichment of histone H3 phosphorylation on 'Ser-10' (H3S10ph) in LPS-stimulated myeloid cells. This phosphorylation enhances the accessibility of the cryptic NF-kappa-B-binding sites marking promoters for increased NF-kappa-B recruitment. Phosphorylates CDC25B and CDC25C which is required for binding to 14-3-3 proteins and leads to initiation of a G2 delay after ultraviolet radiation. Phosphorylates TIAR following DNA damage, releasing TIAR from GADD45A mRNA and preventing mRNA degradation. The p38 MAPKs may also have kinase-independent roles, which are thought to be due to the binding to targets in the absence of phosphorylation. Protein O-Glc-N-acylation catalyzed by the OGT is regulated by MAPK14, and, although OGT does not seem to be phosphorylated by MAPK14, their interaction increases upon MAPK14 activation induced by glucose deprivation. This interaction may regulate OGT activity by recruiting it to specific targets such as neurofilament H, stimulating its O-Glc-N-acylation. Required in mid-fetal development for the growth of embryo-derived blood vessels in the labyrinth layer of the placenta. Also plays an essential role in developmental and stress-induced erythropoiesis, through regulation of EPO gene expression. Phosphorylates S100A9 at 'Thr-113' (By similarity).
• Uniprot ID: P70618
• Protein Accession #: NP_112282.2
• Nucleotide Accession #: NM_031020.2
• Protein Size (# AA): 360