Phospho-eIF4E (S209) and Total eIF4E ELISA Kit (ab279777)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phospho-eIF4E (S209) and Total eIF4E ELISA Kit
See all eIF4E kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phospho-eIF4E (S209) and Total eIF4E (ab279777) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated eIF4E protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-eIF4E and total eIF4E. An anti-pan eIF4E antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and eIF4E present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-eIF4E (S209) antibody is added to detect phosphorylated eIF4E. In the remaining wells, rabbit anti-pan-eIF4E antibody is used to detect pan eIF4E. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of eIF4E (S209) or pan eIF4E bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer Concentrate 1 x 25ml 2X Cell lysate buffer 1 x 10ml 5X Assay Diluent 1 x 15ml HRP-conjugated anti-rabbit IgG concentrate (1000X) 1 vial Pan-eIF4E Coated Microplate 1 unit Positive Control - treated HeLa cell lysate 1 vial Rabbit anti-phospho-eIF4E (S209)-antibody 1 vial Rabbit anti-pan-eIF4E detection antibody 1 vial Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
Research areas
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Function
Its translation stimulation activity is repressed by binding to the complex CYFIP1-FMR1 (By similarity). 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. Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression. In the CYFIP1-EIF4E-FMR1 complex this subunit mediates the binding to the mRNA cap. -
Sequence similarities
Belongs to the eukaryotic initiation factor 4E family. -
Post-translational
modificationsPhosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex. - Information by UniProt
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Alternative names
- AUTS19
- CBP
- eIF 4E
see all -
Database links
- Entrez Gene: 1977 Human
- Omim: 133440 Human
- SwissProt: P06730 Human
- Unigene: 249718 Human
Images
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Positive Control
HeLa cells were treated with Anisomycin.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
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HeLa cells treated with/without Anisomycin.HeLa cells were untreated or treated with Anisomycin. -
HeLa cells treated with/without Anisomycin.HeLa cells were untreated or treated with Anisomycin.