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. The medium was removed and cells were lysed with 4 mL 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. Soluble supernatants were collected after a 10 min centrifugation, pre-diluted and tested using the two-plate assay protocol.)
4EBP1 assay kit
Phospho-4EBP1 (T37/T46) TR-FRET Cellular Assay Kit
Acc-phospho-4EBP1 (20 uL)
4EBP1 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. The medium was removed and cells were lysed with 4 mL 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. Soluble supernatants were collected after a 10 min centrifugation, pre-diluted 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. The medium was removed and cells were lysed with 4 mL 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. Soluble supernatants were collected after a 10 min centrifugation, pre-diluted and tested using the two-plate assay protocol.)
Testing Data
(A431 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 with PP242 for 3 h at 37 degree C. 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 phospho-4EBP1 (T37/T46) 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
(A431 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 with PP242 for 3 h at 37 degree C. 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 phospho-4EBP1 (T37/T46) 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
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• Signal stable over time :: Flexibility to read plate
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• 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
This gene encodes one member of a family of translation repressor proteins. The protein directly interacts with eukaryotic translation initiation factor 4E (eIF4E), which is a limiting component of the multisubunit complex that recruits 40S ribosomal subunits to the 5' end of mRNAs. Interaction of this protein with eIF4E inhibits complex assembly and represses translation. This protein is phosphorylated in response to various signals including UV irradiation and insulin signaling, resulting in its dissociation from eIF4E and activation of mRNA translation. [provided by RefSeq, Jul 2008]
Uniprot Description
4E-BP1: binds to eIF4E, preventing its assembly into the EIF4F complex and inhibiting cap-dependent translation. Phosphorylation of 4E-BP1 disrupts this binding, activating cap-dependent translation. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the PI3 kinase pathway.
Protein type: Translation; Translation initiation
Chromosomal Location of Human Ortholog: 8p12
Cellular Component: nucleoplasm; protein complex; cytoplasm; cytosol
Molecular Function: protein binding; translation repressor activity; eukaryotic initiation factor 4E binding
Biological Process: negative regulation of translational initiation; response to ethanol; cellular protein metabolic process; TOR signaling pathway; translation; insulin receptor signaling pathway; translational initiation; gene expression; positive regulation of mitotic cell cycle; negative regulation of protein complex assembly; G1/S transition of mitotic cell cycle; lung development
Research Articles on 4EBP1
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Product Notes
The Human, Mouse, Rat, Monkey 4EBP1 eif4ebp1 (Catalog #AAA298001) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298001 Assay Kit recognizes Human, Mouse, Rat, Monkey 4EBP1. AAA Biotech's 4EBP1 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 4EBP1 eif4ebp1 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 "4EBP1, 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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