Control Lysate
(HEK293 cells were grown in a T175 flask at 37 degree C, 5%CO2, in EMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #1 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 15 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
ERK1/2 assay kit
Total ERK1/2 TR-FRET Cellular Assay Kit
Acc-Total ERK (20 uL)
ERK control lysate (100 uL)
5X Lysis Buffer #1 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(HEK293 cells were grown in a T175 flask at 37 degree C, 5%CO2, in EMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #1 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 15 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(HEK293 cells were grown in a T175 flask at 37 degree C, 5%CO2, in EMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 30 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #1 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 15 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(HEK293 cells were seeded at 50,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS. Cells were then treated with either EGF for 15 min (stimulation) or with erlotinib for 15 min then EGF (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #1 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 15 min at room temperature. The lysates were then analyzed for total-ERK1/2 using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-ERK1/2 (T202/Y204) are also shown for comparison. 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
(HEK293 cells were seeded at 50,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS. Cells were then treated with either EGF for 15 min (stimulation) or with erlotinib for 15 min then EGF (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #1 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 15 min at room temperature. The lysates were then analyzed for total-ERK1/2 using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-ERK1/2 (T202/Y204) are also shown for comparison. 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 MAP kinase family. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act in a signaling cascade that regulates various cellular processes such as proliferation, differentiation, and cell cycle progression in response to a variety of extracellular signals. This kinase is activated by upstream kinases, resulting in its translocation to the nucleus where it phosphorylates nuclear targets. Alternatively spliced transcript variants encoding different protein isoforms have been described. [provided by RefSeq, Jul 2008]
Uniprot Description
ERK1: a serine/threonine kinase of the GMGC group that plays a critical role in the regulation of cell growth and differentiation. ERK1 (MAPK3) and ERK2 (MAPK1) play central roles in MAPK cascades and are activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones and neurotransmitters. Depending on the cellular context, MAPK cascades mediate diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. MAPK cascades also plays a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. Activation of MAP kinases occurs through phosphorylation of threonine and tyrosine residues at the sequence T*EY* by upstream MAP kinase kinases, MEK1 and -2. Phosphorylation of both the threonine and tyrosine are required for activity. This phosphorylation causes dramatic conformational changes, which enable full activation and interaction of MAPK1/ERK2 with its substrates.
Protein type: Protein kinase, CMGC; Protein kinase, Ser/Thr (non-receptor); EC 2.7.11.24; Kinase, protein; CMGC group; MAPK family; ERK subfamily; MAPK/ERK subfamily
Chromosomal Location of Human Ortholog: 16p11.2
Cellular Component: Golgi apparatus; focal adhesion; mitochondrion; early endosome; nuclear envelope; pseudopodium; caveola; cytosol; nucleoplasm; microtubule cytoskeleton; cytoskeleton; late endosome; nucleus
Molecular Function: MAP kinase activity; protein binding; phosphotyrosine binding; ATP binding; phosphatase binding
Biological Process: axon guidance; viral reproduction; nerve growth factor receptor signaling pathway; positive regulation of histone phosphorylation; DNA damage induced protein phosphorylation; activation of MAPKK activity; apoptosis; activation of MAPK activity; stress-activated MAPK cascade; toll-like receptor 3 signaling pathway; sensory perception of pain; protein amino acid phosphorylation; toll-like receptor 10 signaling pathway; BMP signaling pathway; toll-like receptor 5 signaling pathway; response to exogenous dsRNA; regulation of transcription factor activity; small GTPase mediated signal transduction; lipopolysaccharide-mediated signaling pathway; toll-like receptor 4 signaling pathway; epidermal growth factor receptor signaling pathway; platelet activation; fibroblast growth factor receptor signaling pathway; MyD88-independent toll-like receptor signaling pathway; cytokine and chemokine mediated signaling pathway; MAPKKK cascade; transcription from RNA polymerase I promoter; cell cycle; toll-like receptor 2 signaling pathway; MyD88-dependent toll-like receptor signaling pathway; regulation of stress-activated MAPK cascade; organ morphogenesis; cartilage development; Ras protein signal transduction; toll-like receptor signaling pathway; insulin receptor signaling pathway; innate immune response; positive regulation of transcription from RNA polymerase II promoter; positive regulation of histone acetylation; gene expression; positive regulation of protein amino acid phosphorylation; toll-like receptor 9 signaling pathway; vascular endothelial growth factor receptor signaling pathway; blood coagulation; transcription initiation from RNA polymerase I promoter; phosphorylation; regulation of cytoskeleton organization and biogenesis
Research Articles on ERK1/2
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Product Notes
The Human, Mouse, Rat, Monkey, Mink, Zebrafish, Bovine, Pig ERK1/2 mapk3 (Catalog #AAA298024) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298024 Assay Kit recognizes Human, Mouse, Rat, Monkey, Mink, Zebrafish, Bovine, Pig ERK1/2. AAA Biotech's ERK1/2 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 ERK1/2 mapk3 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 "ERK1/2, 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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