Control Lysate
(A431 cells were grown in a T175 flask at 37 degree C/5% CO2 in DMEM with 10% FBS and cultured to 70% confluence. Stimulation was done with 1 nM EGF for 10 min at 37oC. After removal of cell culture medium, 4 mL of 1X Lysis Buffer #2 supplemented with 1 mM NaF and 2 mM Na3VO4 were added and incubated for 30 min. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
CREB assay kit
Total CREB TR-FRET Cellular Assay Kit
Predicted: Rat
Predicted: Rat
Acc-Total CREB (20 uL)
CREB control lysate (100 uL)
5X Lysis Buffer #2 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(A431 cells were grown in a T175 flask at 37 degree C/5% CO2 in DMEM with 10% FBS and cultured to 70% confluence. Stimulation was done with 1 nM EGF for 10 min at 37oC. After removal of cell culture medium, 4 mL of 1X Lysis Buffer #2 supplemented with 1 mM NaF and 2 mM Na3VO4 were added and incubated for 30 min. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(A431 cells were grown in a T175 flask at 37 degree C/5% CO2 in DMEM with 10% FBS and cultured to 70% confluence. Stimulation was done with 1 nM EGF for 10 min at 37oC. After removal of cell culture medium, 4 mL of 1X Lysis Buffer #2 supplemented with 1 mM NaF and 2 mM Na3VO4 were added and incubated for 30 min. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(Cells were seeded at 50,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were then treated at 37oC with either forskolin for 30 min (stimulation; NIH 3T3) or erlotinib for 30 min then 1 nM EGF for 10 min (inhibition; A431). 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 for total-CREB using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-CREB (S133) 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
(Cells were seeded at 50,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were then treated at 37oC with either forskolin for 30 min (stimulation; NIH 3T3) or erlotinib for 30 min then 1 nM EGF for 10 min (inhibition; A431). 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 for total-CREB using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-CREB (S133) 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
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• TR-FRET technology :: Proven, sensitive and robust assay technology
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NCBI and Uniprot Product Information
NCBI Description
This gene encodes a transcription factor that is a member of the leucine zipper family of DNA binding proteins. This protein binds as a homodimer to the cAMP-responsive element, an octameric palindrome. The protein is phosphorylated by several protein kinases, and induces transcription of genes in response to hormonal stimulation of the cAMP pathway. Alternate splicing of this gene results in several transcript variants encoding different isoforms. [provided by RefSeq, Mar 2016]
Uniprot Description
CREB: a transcription factor of the leucine zipper family of DNA binding proteins. Binds as a homodimer to the cAMP-responsive element (CRE). Phosphorylated by several protein kinases, and induces transcription of genes in response to hormonal stimulation of the cAMP pathway. Two splice-variant isoforms have been described.
Protein type: Transcription factor; DNA-binding
Chromosomal Location of Human Ortholog: 2q34
Cellular Component: nucleoplasm; nucleus
Molecular Function: RNA polymerase II transcription factor activity, enhancer binding; protein binding; enzyme binding; transcription cofactor activity; transcription factor activity
Biological Process: transcription from RNA polymerase II promoter; circadian rhythm; lactation; axon guidance; response to glucagon stimulus; viral reproduction; nerve growth factor receptor signaling pathway; positive regulation of osteoclast differentiation; positive regulation of transcription, DNA-dependent; stress-activated MAPK cascade; positive regulation of multicellular organism growth; toll-like receptor 3 signaling pathway; signal transduction; protein amino acid phosphorylation; toll-like receptor 10 signaling pathway; response to organic substance; toll-like receptor 5 signaling pathway; synaptic transmission; toll-like receptor 4 signaling pathway; response to drug; epidermal growth factor receptor signaling pathway; mitochondrion organization and biogenesis; Notch signaling pathway; fibroblast growth factor receptor signaling pathway; phosphoinositide-mediated signaling; protein stabilization; secretory granule organization and biogenesis; MyD88-independent toll-like receptor signaling pathway; organelle organization and biogenesis; positive regulation of hormone secretion; positive regulation of lipid biosynthetic process; memory; toll-like receptor 2 signaling pathway; MyD88-dependent toll-like receptor signaling pathway; phospholipase C activation; pituitary gland development; toll-like receptor signaling pathway; positive regulation of fat cell differentiation; innate immune response; positive regulation of transcription from RNA polymerase II promoter; toll-like receptor 9 signaling pathway; regulation of cell size
Disease: Histiocytoma, Angiomatoid Fibrous
Research Articles on CREB
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Product Notes
The Human, Mouse Predicted: Rat CREB creb1 (Catalog #AAA298012) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298012 Assay Kit recognizes Human, Mouse Predicted: Rat CREB. AAA Biotech's CREB 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 CREB creb1 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 "CREB, 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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