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Anti-EIF4E Antibody Y448
Also for EIF4E (NM_001968)
|A synthetic peptide corresponding to residues in human eIF-4E, was used as immunogen. The antibody detects a band on western blot of approximately 28 kDa.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:500; IHC-P: Use at an assay dependent dilution; ICC: 1:250 - 1:500; FC: Use 1?g for 106<:sup> cells; IP: 1:30
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Homo sapiens eukaryotic translation initiation factor 4E (EIF4E), transcript variant 1|
|AUTS19; CBP; EIF4E1; EIF4EL1; EIF4F|
|eIF-4E is a eukaryotic translation initiation factor involved in directing ribosomes to the cap structure of mRNAs. It exists in two forms: as a free form (25 kDa) and as part of a multiprotein complex eIF-4F (1). eIF-4E appears to be the least abundant of the initiation factors and acts as a rate-limiting step of initiation (2). Since translation is regulated by phosphorylation, eIF-4E phosphorylation at Ser 209 by MAPK signal-integrating kinase 1 (Mnk1) and kinase 2 (Mnk2) may directly regulate the rate of protein synthesis initiation (3). There is also evidence that eIF-4E can function as an oncogene (4-5). |
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Western blot - eIF4E antibody [Y448]; Anti-eIF4E antibody [Y448] at 1/500 dilution + 293 cell lysate.Predicted band size : 25 kDa.Observed band size : 30 kDa .
Immunohistochemistry (Paraffin-embedded sections) - eIF4E antibody [Y448]; This image shows human breast carcinoma stained with ab33766
Immunocytochemistry/ Immunofluorescence - Anti-eIF4E antibody [Y448]; ICC/IF image of ab33766 stained MCF7 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry - Anti-eIF4E antibody [Y448]; Overlay histogram showing HEK293 cells stained with ab33766 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (1µg/1x10^6 cells ) used under the same conditions. Acquisition of >5,000 events was performed.