> Last Item Added:
0 Items

Ordering Information

  • : 17-487
  • : 96 assays
  • Product Family Information

Product Images

Typical p38α Standard Curve 100 αL of progressive 2 fold dilutions of the p38α standard included in the kit and run as described in the assay instruct...

p38α STAR ELISA Kit

Description:
p38α STAR ELISA Kit
Trade Name:
Upstate (Millipore)
Qty/Pk:
96 assays
Background Information:
I. TEST PRINCIPLE
The 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 p38alpha plate is coated with a specific mouse monoclonal p38alpha capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standards included in the kit are incubated in the microwells allowing p38alpha to be captured in the plate wells. The plate is then washed to remove any unbound non-specific material. A specific rabbit anti-p38 MAPK antibody is added to detect the captured p38α 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. This allows for a sensitive enzymatic detection of the sample.
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 generate a standard curve.

II. p38 MAPK 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 latter p38 isoforms share 74%, 57%, and 60% sequence homology 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 and transcription factors including STAT1, p53, NFATp, ATF-2, Elk-1 (although weakly), and MEF2a/c. The 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 is.
Applications:
This p38α STAR ELISA Kit is used to measure & quantify p38/SAPK levels.
Key Applications:
ELISA
Application Notes:
• All kit reagents should be brought to room temperature (20°C to 25°C) prior to use.
• Do not use reagents beyond the expiration date of the kit.
• Do not mix or interchange reagent from various kit lots.
• Manual Plate Washing: Vigorous washing and complete removal of all liquid by aspiration at the end of each washing step is very important to obtain low background values. Gentle agitation during the wash steps or a 2-3 minute soak may reduce background values.
• The desiccant enclosed in the 96-well capture plate pouch will keep the plate stable when stored at 2° to 8°C should the pouch loose its seal during shipping.
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Entrez Gene Summary:
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. This kinase is activated by various environmental stresses and proinflammatory cytokines. The activation requires its phosphorylation by MAP kinase kinases (MKKs), or its autophosphorylation triggered by the interaction of MAP3K7IP1/TAB1 protein with this kinase. The substrates of this kinase include transcription regulator ATF2, MEF2C, and MAX, cell cycle regulator CDC25B, and tumor suppressor p53, which suggest the roles of this kinase in stress related transcription and cell cycle regulation, as well as in genotoxic stress response. Four alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported.
UniProt Summary:
FUNCTION: SwissProt: Q16539 # Responds 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.
COFACTOR: Magnesium.
SIZE: 360 amino acids; 41293 Da
SUBUNIT: Binds to a kinase interaction motif within the protein tyrosine phosphatase, PTPRR. This interaction retains MAPK14 in the cytoplasm and prevents nuclear accumulation. Interacts with SPAG9 (By similarity). Interacts with NP60.
SUBCELLULAR LOCATION: Cytoplasm (By similarity). Nucleus (By similarity).
TISSUE SPECIFICITY: Brain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.DOMAIN:SwissProt: Q16539 The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
PTM: Dually phosphorylated on Thr-180 and Tyr-182, which activates the enzyme. & Phosphorylated upon DNA damage, probably by ATM or ATR.
SIMILARITY: Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily. & Contains 1 protein kinase domain.
Brand Family:
Upstate
Sensitivity:
Sensitivity: 15 pg/mL
Range of Detection: 31.25 to 2000 pg/mL
Storage Conditions:
Maintain the unopened kit at 2-8°C until expiration date.
Gene Symbol:
  • MAPK14
  • PRKM14
  • RK
  • SAPK2A
  • CSBP1
  • p38
  • CSPB1
  • CSBP
  • PRKM15
  • Mxi2
  • p38ALPHA
  • MXI2
  • EXIP
  • CSBP2
Product Name:
p38α STAR ELISA Kit
Materials Required but Not Delivered:
Materials Not Supplied 1. Multi-channel or repeating pipettes 2. Plate shaker (optional) 3. Pipettors and tips capable of accurately measuring 1-1000 µL 4. Graduated serological pipettes 5. 96-well microtiter Plate Reader with 450 nm filter 6. Graphing software for plotting data or graph paper for manual plotting of data 7. Microfuge tubes for standard and sample dilutions 8. Mechanical vortex 9. 1 liter container 10. Distilled or deionized water
Analytes Available:
p38/SAPK
Components:
  • Capture Plate pre-coated with anti-p38alpha antibody: (Part No. 17-487A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.
  • Anti-p38 detection antibody: (Part No. 17-487B) One bottle (11 mL) of anti- p38 MAPK detection antibody containing sodium azide, ready to use.
  • ELISA Diluent: (Part No. 17-487C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.
  • 25X ELISA Wash Buffer: (Part No. 17-487D) One bottle (50 mL) of 25X ELISA Wash Buffer.
  • Anti-Rabbit IgG HRP conjugate: (Part No. 17-487E) One vial (125 µL) of 100X anti-rabbit HRP conjugate containing thimerosal.
  • HRP Diluent: (Part No. 17-487F) One bottle (25 mL) of HRP Diluent containing thimerosal.
  • TMB Solution: (Part No. 17-487G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use.
  • Stop Solution: (Part No. 17-487H) One bottle (25 mL) of stop solution, ready to use.
  • p38 MAPK Standard: (Part No. 17-487I) Two vials of p38 standard, lyophilized.
  • Plate Covers: Two plate covers.
UniProt Number:
Detection Methods:
Colorimetric
Entrez Gene Number: