Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5% CO2 in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM PMA for 5 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
MEK1 assay kit
Phospho-MEK1 (S218/222) TR-FRET Cellular Assay Kit
Acc-phospho-MEK1 (20 uL)
MEK1 control lysate (100 uL)
5X Lysis Buffer #3 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5% CO2 in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM PMA for 5 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5% CO2 in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM PMA for 5 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(HeLa cells were seeded at 25,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were then treated at 37 degree C with either PMA for 5 min (stimulation) or L779,450 for 15 min then 30 uM PMA (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #3 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-MEK1 (Ser218/222) using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 4 h incubation period at room temperature.)
Testing Data
(HeLa cells were seeded at 25,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were then treated at 37 degree C with either PMA for 5 min (stimulation) or L779,450 for 15 min then 30 uM PMA (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #3 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-MEK1 (Ser218/222) using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 4 h incubation period at room temperature.)
A superior alternative to ELISA
Features :: Benefits
• TR-FRET technology :: Proven, sensitive and robust assay technology
• Homogeneous, add-and-read assay :: No wash, no separation steps
• < 1 hour bench time :: Faster, easier than ELISA
• Adherent or suspension cells :: Higher flexibility
• Validated assays :: Results you can trust
• Small sample volume (15uL) :: Save precious samples
• Compatible with low-volume 96-well and 384-well plates : Higher throughput
• Signal stable over time :: Flexibility to read plate
• Kits available in 100 and 500 assay points :: Cost-effective and flexible
• Kits available for phospho, total and phospho+total protein :: Allows multiple protein measurements in one plate from one sample
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is a member of the dual specificity protein kinase family, which acts as a mitogen-activated protein (MAP) kinase kinase. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act as an integration point for multiple biochemical signals. This protein kinase lies upstream of MAP kinases and stimulates the enzymatic activity of MAP kinases upon wide variety of extra- and intracellular signals. As an essential component of MAP kinase signal transduction pathway, this kinase is involved in many cellular processes such as proliferation, differentiation, transcription regulation and development. [provided by RefSeq, Jul 2008]
Uniprot Description
MEK1: a dual-specificity protein kinase of the STE7 kinase family. Phosphorylated and activated by Raf, Mos and Cot kinases. Phosphorylates a Thr and a Tyr residue in a Thr-Glu-Tyr sequence located in the activation loop of ERK1 and ERK2. An essential component of MAP kinase signal transduction pathways involved in many cellular processes such as proliferation, differentiation, transcription regulation and development.
Protein type: Protein kinase, STE; Protein kinase, dual-specificity (non-receptor); EC 2.7.12.2; Kinase, protein; STE group; STE7 family
Chromosomal Location of Human Ortholog: 15q22.1-q22.33
Cellular Component: dendrite cytoplasm; Golgi apparatus; microtubule; focal adhesion; mitochondrion; endoplasmic reticulum; early endosome; perikaryon; cell cortex; cytosol; axon; perinuclear region of cytoplasm; cytoplasm; late endosome; plasma membrane; microtubule organizing center; nucleus
Molecular Function: MAP kinase kinase activity; protein serine/threonine kinase activity; protein serine/threonine kinase activator activity; protein binding; Ras GTPase binding; receptor signaling protein tyrosine phosphatase activity; protein-tyrosine kinase activity; protein serine/threonine/tyrosine kinase activity; mitogen-activated protein kinase kinase kinase binding; ATP binding; protein kinase activity
Biological Process: axon guidance; peptidyl-tyrosine phosphorylation; activation of MAPKK activity; nerve growth factor receptor signaling pathway; protein heterooligomerization; activation of MAPK activity; negative regulation of homotypic cell-cell adhesion; response to glucocorticoid stimulus; stress-activated MAPK cascade; cell motility involved in cell locomotion; pathogenesis; toll-like receptor 3 signaling pathway; chemotaxis; signal transduction; toll-like receptor 10 signaling pathway; neuron differentiation; toll-like receptor 5 signaling pathway; negative regulation of cell proliferation; positive regulation of RNA elongation from RNA polymerase II promoter; small GTPase mediated signal transduction; vesicle transport along microtubule; response to axon injury; cell cycle arrest; toll-like receptor 4 signaling pathway; Golgi inheritance; epidermal growth factor receptor signaling pathway; mitosis; fibroblast growth factor receptor signaling pathway; MyD88-independent toll-like receptor signaling pathway; MAPKKK cascade; melanosome transport; toll-like receptor 2 signaling pathway; MyD88-dependent toll-like receptor signaling pathway; keratinocyte differentiation; regulation of stress-activated MAPK cascade; cell proliferation; regulation of vascular smooth muscle contraction; Ras protein signal transduction; insulin receptor signaling pathway; toll-like receptor signaling pathway; innate immune response; positive regulation of Ras protein signal transduction; toll-like receptor 9 signaling pathway; response to oxidative stress; cell motility; positive regulation of cell differentiation; vascular endothelial growth factor receptor signaling pathway; positive regulation of cell migration
Disease: Noonan Syndrome 1; Cardiofaciocutaneous Syndrome 3
Research Articles on MEK1
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Product Notes
The Human, Mouse, Rat, Monkey MEK1 map2k1 (Catalog #AAA298033) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298033 Assay Kit recognizes Human, Mouse, Rat, Monkey MEK1. AAA Biotech's MEK1 can be used in a range of immunoassay formats including, but not limited to, Cell-Based Assays. Researchers should empirically determine the suitability of the MEK1 map2k1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "MEK1, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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