Control Lysate
(HEK293cells 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 1 uM of anisomycin for 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2containing 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.)
JNK1/2/3 assay kit
Phospho-JNK1/2/3 (T183/Y185) + Total JNK1/2/3 TR-FRET Cellular Assay Kit
Acc-phospho-JNK (80 uL)
Eu-Total JNK (5 uL)
Acc-Total JNK (20 uL)
JNK control lysate (500 uL)
5X Lysis Buffer #2 (5 mL)
10X TR-FRET Detection buffer (250 uL)
Control Lysate
(HEK293cells 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 1 uM of anisomycin for 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2containing 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
(HEK293cells 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 1 uM of anisomycin for 60 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2containing 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
(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 treated with either anisomycin for 60 min (stimulation) or BX 795 for 30 min then anisomycin (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 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 on separate wells for phospho-JNK1/2/3 (T183/Y185) and total-JNK1/2/3 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
(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 treated with either anisomycin for 60 min (stimulation) or BX 795 for 30 min then anisomycin (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 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 on separate wells for phospho-JNK1/2/3 (T183/Y185) and total-JNK1/2/3 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 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 cell stimuli, and targets specific transcription factors, and thus mediates immediate-early gene expression in response to cell stimuli. The activation of this kinase by tumor-necrosis factor alpha (TNF-alpha) is found to be required for TNF-alpha induced apoptosis. This kinase is also involved in UV radiation induced apoptosis, which is thought to be related to cytochrom c-mediated cell death pathway. Studies of the mouse counterpart of this gene suggested that this kinase play a key role in T cell proliferation, apoptosis and differentiation. Several alternatively spliced transcript variants encoding distinct isoforms have been reported. [provided by RefSeq, Apr 2016]
Uniprot Description
JNK1: a protein kinase of the MAPK family that is potently activated by a variety of environmental stresses, including UV and gamma radiation, ceramides, pro-inflammatory cytokines and, in some instances, by growth factors and GPCR agonists. Substrates include a number of transcription factors, primarily components of AP-1 such as c-Jun and ATF2, thus regulating AP-1 transcriptional activity. In T- cells, JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells. Binds to at least four scaffolding proteins, JIP-1, -2, -3 and -4. Activity increased in obesity. Inhibition or mouse knockout increases insulin sensitivity. Part of NFB pathway involved in inflammation and cancer, and signals downstream of Ras, though possibly as an apoptotic negative regulator of growth. Four alternatively-spliced isoforms have been described.
Protein type: Kinase, protein; EC 2.7.11.24; Protein kinase, CMGC; Protein kinase, Ser/Thr (non-receptor); CMGC group; MAPK family; MAPK/JNK subfamily; JNK subfamily
Chromosomal Location of Human Ortholog: 10q11.22
Cellular Component: nucleoplasm; mitochondrion; cytosol; nucleus
Molecular Function: protein serine/threonine kinase activity; protein binding; enzyme binding; histone deacetylase regulator activity; histone deacetylase binding; JUN kinase activity; ATP binding
Biological Process: nerve growth factor receptor signaling pathway; apoptosis; positive regulation of apoptosis; rhythmic process; stress-activated MAPK cascade; toll-like receptor 3 signaling pathway; protein amino acid phosphorylation; toll-like receptor 10 signaling pathway; toll-like receptor 5 signaling pathway; regulation of protein localization; regulation of transcription factor activity; response to stress; JNK cascade; negative regulation of protein binding; toll-like receptor 4 signaling pathway; response to UV; ossification; regulation of histone deacetylation; MyD88-independent toll-like receptor signaling pathway; peptidyl-threonine phosphorylation; regulation of circadian rhythm; JUN phosphorylation; toll-like receptor 2 signaling pathway; MyD88-dependent toll-like receptor signaling pathway; peptidyl-serine phosphorylation; response to cadmium ion; toll-like receptor signaling pathway; innate immune response; toll-like receptor 9 signaling pathway; negative regulation of apoptosis
Research Articles on JNK1/2/3
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Product Notes
The Human, Mouse, Rat JNK1/2/3 mapk8 (Catalog #AAA298031) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298031 Assay Kit recognizes Human, Mouse, Rat JNK1/2/3. AAA Biotech's JNK1/2/3 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 JNK1/2/3 mapk8 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 "JNK1/2/3, 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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