Control Lysate
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2 in EMEM with 10% FBS, 10% non-essential amino-acids and cultured to 80% confluence. The medium was removed and cells were stimulated with 3 uM Insulin for 30 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3supplemented 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.)
GSK3beta assay kit
Total GSK3beta TR-FRET Cellular Assay Kit
Acc-Total GSK3b (20 uL)
GSK3b control lysate (100 uL)
5X Lysis Buffer #3 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2 in EMEM with 10% FBS, 10% non-essential amino-acids and cultured to 80% confluence. The medium was removed and cells were stimulated with 3 uM Insulin for 30 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3supplemented 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
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2 in EMEM with 10% FBS, 10% non-essential amino-acids and cultured to 80% confluence. The medium was removed and cells were stimulated with 3 uM Insulin for 30 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3supplemented 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
(MCF-7 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS and 10% non-essential amino-acids. Cells were then treated at room temperature with either Insulin for 30 min (stimulation) or UCN-01 for 2 hours then 100 nM Insulin for 30 min (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 total-GSK3betausing the two-plate assay protocol.Lysates analyzed on separate wells for phospho-GSK3beta (Ser9) are also shown for comparison. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 2 h incubation period at room temperature.)
Testing Data
(MCF-7 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS and 10% non-essential amino-acids. Cells were then treated at room temperature with either Insulin for 30 min (stimulation) or UCN-01 for 2 hours then 100 nM Insulin for 30 min (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 total-GSK3betausing the two-plate assay protocol.Lysates analyzed on separate wells for phospho-GSK3beta (Ser9) are also shown for comparison. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 2 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 serine-threonine kinase belonging to the glycogen synthase kinase subfamily. It is a negative regulator of glucose homeostasis and is involved in energy metabolism, inflammation, ER-stress, mitochondrial dysfunction, and apoptotic pathways. Defects in this gene have been associated with Parkinson disease and Alzheimer disease. [provided by RefSeq, Aug 2017]
Uniprot Description
GSK3B: a proline-directed protein kinase of the GSK family. Phosphorylates and inactivates glycogen synthase. Participates in the Wnt signaling pathway. Involved in energy metabolism, neuronal cell development, and body pattern formation
Protein type: Protein kinase, CMGC; Kinase, protein; Protein kinase, Ser/Thr (non-receptor); EC 2.7.11.1; EC 2.7.11.26; CMGC group; GSK family; GSK subfamily
Chromosomal Location of Human Ortholog: 3q13.3
Cellular Component: centrosome; dendritic spine; cytosol; beta-catenin destruction complex; ribonucleoprotein complex; lipid raft; nucleoplasm; growth cone; cell soma; perinuclear region of cytoplasm; cytoplasm; plasma membrane; dendritic shaft; nucleus
Molecular Function: p53 binding; beta-catenin binding; protein kinase binding; integrin binding; ionotropic glutamate receptor binding; protein serine/threonine kinase activity; NF-kappaB binding; protein binding; ubiquitin protein ligase binding; tau-protein kinase activity; kinase activity; tau protein binding; ATP binding
Biological Process: circadian rhythm; glycogen metabolic process; fat cell differentiation; axon guidance; negative regulation of MAP kinase activity; genetic imprinting; re-entry into mitotic cell cycle; nerve growth factor receptor signaling pathway; positive regulation of protein binding; protein amino acid autophosphorylation; response to lithium ion; positive regulation of axon extension; Wnt receptor signaling pathway through beta-catenin; protein amino acid phosphorylation; positive regulation of protein export from nucleus; protein export from nucleus; negative regulation of protein binding; positive regulation of cell-matrix adhesion; ER overload response; myoblast fusion; negative regulation of glycogen biosynthetic process; epidermal growth factor receptor signaling pathway; response to drug; regulation of microtubule-based process; cell migration; phosphoinositide-mediated signaling; fibroblast growth factor receptor signaling pathway; hippocampus development; negative regulation of dendrite morphogenesis; negative regulation of NFAT protein import into nucleus; positive regulation of peptidyl-serine phosphorylation; organ morphogenesis; negative regulation of neuron maturation; peptidyl-serine phosphorylation; positive regulation of protein complex assembly; establishment of cell polarity; positive regulation of proteasomal ubiquitin-dependent protein catabolic process; positive regulation of protein catabolic process; epithelial to mesenchymal transition; innate immune response; positive regulation of transcription from RNA polymerase II promoter; negative regulation of protein complex assembly; regulation of neuronal synaptic plasticity; negative regulation of apoptosis
Research Articles on GSK3beta
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Product Notes
The Human, Mouse, Rat GSK3beta gsk3b (Catalog #AAA298028) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298028 Assay Kit recognizes Human, Mouse, Rat GSK3beta. AAA Biotech's GSK3beta 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 GSK3beta gsk3b 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 "GSK3beta, 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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