Control Lysate
(HEK293 cells were grown in a T175 flask at 37 degree C, 5% CO2 in EMEM with 10% FBS and cultured to 85% confluence. Stimulation was done with 33 nM EGF for 30 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2supplemented 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.)
eIF4E assay kit
Phospho-eIF4E (S209) TR-FRET Cellular Assay Kit
Acc-phospho-eIF4E (20 uL)
eIF4E control lysate (100 uL)
5X Lysis Buffer #2 (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 85% confluence. Stimulation was done with 33 nM EGF for 30 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2supplemented 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
(HEK293 cells were grown in a T175 flask at 37 degree C, 5% CO2 in EMEM with 10% FBS and cultured to 85% confluence. Stimulation was done with 33 nM EGF for 30 min at 37 degree C. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2supplemented 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
(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 CPG 57380 for 30 min at 37 degree C, then 10 nM EGF for 10 min. 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-eIF4E (S209) using the two-plate assay protocol. The TR-FRETsignal 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 CPG 57380 for 30 min at 37 degree C, then 10 nM EGF for 10 min. 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-eIF4E (S209) using the two-plate assay protocol. The TR-FRETsignal 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 component of the eukaryotic translation initiation factor 4F complex, which recognizes the 7-methylguanosine cap structure at the 5' end of messenger RNAs. The encoded protein aids in translation initiation by recruiting ribosomes to the 5'-cap structure. Association of this protein with the 4F complex is the rate-limiting step in translation initiation. This gene acts as a proto-oncogene, and its expression and activation is associated with transformation and tumorigenesis. Several pseudogenes of this gene are found on other chromosomes. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Sep 2015]
Uniprot Description
EIF4E: a protein of the eukaryotic initiation factor 4E family. Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures. Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the EIF4F complex.
Protein type: Translation initiation; Translation
Chromosomal Location of Human Ortholog: 4q23
Cellular Component: mRNA cap complex; stress granule; cytoplasm; eukaryotic translation initiation factor 4F complex; cytosol
Molecular Function: protein binding; eukaryotic initiation factor 4G binding; translation initiation factor activity; RNA cap binding
Biological Process: translation; viral reproduction; cytokine and chemokine mediated signaling pathway; positive regulation of mitotic cell cycle; regulation of translation; poly(A) tail shortening; mRNA export from nucleus; cellular protein metabolic process; insulin receptor signaling pathway; translational initiation; gene expression; mRNA catabolic process, deadenylation-dependent decay; lung development; G1/S transition of mitotic cell cycle
Disease: Autism, Susceptibility To, 19
Research Articles on eIF4E
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Product Notes
The Human, Mouse, Rat eIF4E eif4e (Catalog #AAA298019) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298019 Assay Kit recognizes Human, Mouse, Rat eIF4E. AAA Biotech's eIF4E 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 eIF4E eif4e 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 "eIF4E, 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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