Anti-eIF4E (phospho S209) 抗体 (ab4774)

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Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
  • Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)

Key features and details

  • Rabbit polyclonal to eIF4E (phospho S209)
  • Suitable for: WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-eIF4E (phospho S209) antibody
    eIF4E 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to eIF4E (phospho S209)

  • 由来種

    Rabbit

  • 特異性

    This phosphorylation site specific antibody is selective for eIF-4E containing a phosphate on serine 209.

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Rat, Rabbit, a wide range of other species

  • 免疫原

    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.

  • ポジティブ･コントロール

    • HeLa cell lysate

  • 特記事項

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    pH: 7.30
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 50% Glycerol (glycerin, glycerine), 1% BSA
  • Concentration information loading...

  • 精製度

    Immunogen affinity purified

  • 特記事項（精製）

    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.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab4774 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

ターゲット情報

  • 機能

    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.

  • 配列類似性

    Belongs to the eukaryotic initiation factor 4E family.

  • 翻訳後修飾

    Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • AUTS19 antibody
    • CBP antibody
    • eIF 4E antibody
    • eIF 4F 25 kDa subunit antibody
    • EIF 4F antibody
    • eIF-4E antibody
    • eIF-4F 25 kDa subunit antibody
    • eIF4E antibody
    • EIF4E1 antibody
    • EIF4EL1 antibody
    • EIF4F antibody
    • Eukaryotic translation initiation factor 4 E antibody
    • Eukaryotic translation initiation factor 4E antibody
    • Eukaryotic translation initiation factor 4E like 1 antibody
    • IF4E_HUMAN antibody
    • Messanger RNA Cap Binding Protein eIF 4E antibody
    • MGC111573 antibody
    • mRNA cap binding protein antibody
    • mRNA cap-binding protein antibody
    see all

画像

  • Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
    Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
    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. 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.
  • Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
    Western blot - Anti-eIF4E (phospho S209) antibody (ab4774)
    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 (?) 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 i

参考文献 (1)

ab4774 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4774 は 1 報の論文で使用されています。

  • 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

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Question

What is the concentration of batch 132987?

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Answer

Thank you for your enquiry. The concentration is 0.5 mg/mL. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Here are two blots, one from abCAM and one from cell signaling! And all the details you requested! Lot Number 63956, Abcam order number: 49277, P.O. number: 3P668468. 1. Please describe the problem (high background, wrong band size, more bands, no band etc). Wrong size approximately 36kD and 65kD! 2. On what material are you testing the antibody in WB? Species? Brain tissue lyaste from Guinea Pig along with Pos and Neg cell line lyastae! Cell extract/ Nuclear extract? Whole cell lysate. Purified protein? No Recombinant protein? No 3. How much protein did you load? From 10 to 50 ug, we always run a standard curve with increasing amount of protein to evaluate an antibody of interest before doing the various experimental conditions. How did you prepare the lysate for the analysis (protease inhibito 50 mM Tris-HCl, Standard lysis buffer with protease inhibitors and phosphatase inhibitors. Did you heat the samples? Yes 4. Primary Antibody Specification (in which species was it raised against)? In Rabbit. ab4744, you have all the details! At what dilution(s) have you tested this antibody? 1ug/ml Incubation time, wash step? 2 hours in 1% non-fat dry milk solution in 20 mM TBST, ph 7.6 as suggested in the protocol supplied with the antibody. Three wash steps at room temp for 5 min each. 5. Secondary Antibody Specification (in which species was it raised against)? Goat. Anti-rabbit IgG whole molecule, HRP conjugate At what dilution(s) have you tested this antibody? 1:2500 Incubation time, wash step? 1 hr at room temp, wash step as for primary antibody. Do you know whether the problems you are experiencing come from th No 6. What detection method are you using? Chemiluminescent detection method. 7. Background bands Have you eliminated the possibility that any background bands coul Is the blocking step sufficient? (We recommend blocking the membra YES Are your washing steps sufficiently stringent? (Multiple short was YES At what size are the bands migrating? Could they be degradation pr Approx 36kD and 65kD Please provide an image of your blot (as an e-mail attachment, a f Attached. 8. Optimization attempts How many times have you tried the Western? Two times! And we do western almost everyday in our lab, check the band with your competitor on the image labeled Cell Signaling Technol on the right hand side.. Do you obtain the same results every time e.g. are background band YES What steps have you altered? YES, overnight incubation with primary antibody! 9. Did you apply positive and negative controls along with the samples? Please specify. YES, see band at 36 kD and not at approx. 25kD with abCAM. On the contrary, see band at lower than 29kD with cell signaling tehnol. product. Using abCAM antibody, we did not get band on western blot at the right molecular weight. Please do the needful.

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Answer

Thank you very much for the details that you have provided, and I'm sorry to hear that you are experiencing difficulty with this antibody. As stated on the online datasheet, this particular antibody has been characterized using HeLa cell lysates and has not yet been tested on other species although it is expected to cross-react with a wide range of other species, due to sequence homology. We have not had any other complaints regarding this antibody; the problem may very well be due to the batch you received. I can offer you a replacement batch to try or a refund, please let me know which one you would prefer.

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