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Western blot - eIF4E (phospho S209) antibody (ab4774)
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Western blot - eIF4E (phospho S209) antibody (ab4774)
(enlarge)
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Product Name
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eIF4E (phospho S209) antibody
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See all eIF4E antibodies (10)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to eIF4E (phospho S209)
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Immunogen
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Synthetic peptide (Human) derived from the region of eIF-4E that contains serine 209, based on the Homo sapiens sequence. This region is identical among many species including rat, mouse and rabbit.
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Reacts with
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Hu
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Predicted to react
(species key)
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Ms, Rat, Many, Rb
(Due to sequence homology)
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Specificity
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This phosphorylation site specific antibody is selective for eIF-4E containing a phosphate on serine 209.
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Tested applications
(see key)
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WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
WB: 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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HeLa cell lysate
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Cellular localization
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Cytoplasmic
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Research areas
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Chromatin and Nuclear Signaling >> RNAi >> Eukaryotic Initiation factors (eIF's)
Chromatin and Nuclear Signaling >> DNA / RNA >> RNA Processing >> RNAi
Chromatin and Nuclear Signaling >> DNA / RNA >> Translation >> Regulation
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Relevance
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eIF4E, a protein modulates translation of maternal mRNAs in early embryos before the onset of zygotic transcription. eIF4E also influences the overall rate of translation. eIF4E binds to the 7 methyl GTP cap structure of eukaryotic mRNAs. Phosphorylation of eIF4E on serine 209 regulates the affinity of this protein for the 7 methyl GTP cap and/or RNA. Phosphorylation also enhances the interaction of eIF4E with eIF4G, which form a complex known as eIF4F. eIF4E phosphorylation is correlated with increased translational rate in a number of cell types. Several kinases are currently being investigated as potential regulators of eIF4E including PKC and/or the MAP kinase activated Mnk.
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eIF4E (phospho S209) antibody (ab4774) |
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.1% Sodium Azide
Constituents: 1% BSA, Glycerol, PBS, pH 7.4 Material safety datasheets (MSDS) for this product:
Glycerol MSDS
Sodium Azide MSDS
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Purification notes
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Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated eIF-4E. The final product is generated by affinity chromatography using an eIF-4E-derived peptide that is phosphorylated at serine 209.
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Form
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Liquid
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this eIF4E (phospho S209) antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for eIF4E (phospho S209) antibody images, references, products related to ab4774 and other tools.
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eIF4E (phospho S209) antibody images:
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 Western blot - eIF4E (phospho S209) antibody (ab4774)
Predicted band size : 25 kDa
HeLa cell lysates (A) and HeLa cell lysates treated with alkaline phosphatase (B) were resolved by SDS-PAGE on a 4-20% Tris-glycine gel. The proteins were then transferred to PVDF membrane. Membranes were incubated with 1 µg/mL ab4774. After washing, membranes were incubated with goat (ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method.
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Western blot - eIF4E (phospho S209) antibody (ab4774)
Predicted band size : 25 kDa
Extracts of HeLa cells were resolved by SDS-PAGE on a 4-20% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature, either left untreated (1-4) or treated with lambda (λ) phosphatase (5), and then incubated with the eIF4E (phospho S209) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1,5), the nonphosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Pierce SuperSignal™ method.
The data show that only the peptide corresponding to eIF4E (phospho S209) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
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References for eIF4E (phospho S209) antibody (ab4774)
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This product has been used in:
Kapasi P et al. L13a blocks 48S assembly: role of a general initiation factor in mRNA-specific translational control. Mol Cell 25:113-26 (2007). PubMed: 17218275
If you publish research using ab4774 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to eIF4E
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eIF4E (phospho S209) antibody - more information
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