Anti-EIF2B5 (phospho S539) 抗体 (ab4775)

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Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)
  • Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)

Key features and details

  • Rabbit polyclonal to EIF2B5 (phospho S539)
  • Suitable for: WB
  • Reacts with: Mouse, Rat, Recombinant fragment
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-EIF2B5 (phospho S539) antibody
    EIF2B5 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to EIF2B5 (phospho S539)

  • 由来種

    Rabbit

  • 特異性

    This ab reacts with both human and rat (see, for example, PMID 21165642, 20484009 and 21832246) but it is not very clear if this recognition happens only when S535 (rat) and S544 (human) are phosphorylated or when S535 (rat)/S540 (human) are phosphorylated.

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Mouse, Rat, Recombinant fragment

  • 免疫原

    Synthetic peptide corresponding to Rat EIF2B5.
    (Peptide available as ab5332)

  • ポジティブ･コントロール

    • Recombinant eIF-2B epsilon treated with GSK 3 beta and lambda phosphatase, CHO-T cells transfected with human insulin receptor (IR) +/- insulin. L6, L6 differentiated to myotubes, L6 treated with Insulin (10nM for 10 min), C2C12 and C2C12 treated with Insulin (10nM for 10 min)

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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, 0.1% BSA

    BSA is IgG and protease free
  • 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 pre-adsorbed using (i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated eukaryotic initiation factor 2B epsilon (eIF-2B epsilon), and (ii) a generic serine phosphorylated peptide to remove antibody that is reactive with phospho-serine, irrespective of the sequence. The final product is generated by affinity chromatography using an eIF-2B epsilon-derived peptide that is phosphorylated at serine 539.

  • 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

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab4775の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/1000 - 1/2000. Detects a band of approximately 88 kDa (predicted molecular weight: 82 kDa).
特記事項
WB
1/1000 - 1/2000. Detects a band of approximately 88 kDa (predicted molecular weight: 82 kDa).

ターゲット情報

  • 機能

    Catalyzes the exchange of eukaryotic initiation factor 2-bound GDP for GTP.

  • 関連疾患

    Defects in EIF2B5 are a cause of leukodystrophy with vanishing white matter (VWM) [MIM:603896]. VWM is a leukodystrophy that occurs mainly in children. Neurological signs include progressive cerebellar ataxia, spasticity, inconstant optic atrophy and relatively preserved mental abilities. The disease is chronic-progressive with, in most individuals, additional episodes of rapid deterioration following febrile infections or minor head trauma. While childhood onset is the most common form of the disorder, some severe forms are apparent at birth. A severe, early-onset form seen among the Cree and Chippewayan populations of Quebec and Manitoba is called Cree leukoencephalopathy. Milder forms may not become evident until adolescence or adulthood. Some females with milder forms of the disease who survive to adolescence exhibit ovarian dysfunction. This variant of the disorder is called ovarioleukodystrophy.

  • 配列類似性

    Belongs to the eIF-2B gamma/epsilon subunits family.
    Contains 1 W2 domain.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CACH antibody
      • CLE antibody
      • EI2BE_HUMAN antibody
      • EIF 2B antibody
      • eIF 2B GDP GTP exchange factor antibody
      • eIF 2B GDP GTP exchange factor subunit epsilon antibody
      • eIF-2B GDP-GTP exchange factor subunit epsilon antibody
      • EIF2B antibody
      • EIF2B5 antibody
      • EIF2BE antibody
      • EIF2Bepsilon antibody
      • Eukaryotic Translation Initiation Factor 2 Beta Epsilon antibody
      • Eukaryotic translation initiation factor 2B subunit 5 epsilon 82kDa antibody
      • Eukaryotic translation initiation factor 2B subunit 5 epsilon antibody
      • LVWM antibody
      • Translation initiation factor eIF 2B subunit epsilon antibody
      • Translation initiation factor eIF-2B subunit epsilon antibody
      see all

    画像

    • Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)
      Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)
      Peptide Competition: Recombinant eIF-2B episilon treated with lambda phosphatase (1) or treated with GSK-3 beta (lanes 2-5) and added to background extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.50 µg/mL ab4775 antibody, following prior incubation with: no peptide (1, 2), the nonphosphopeptide corresponding to the immunogen (3), a generic phophoserine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4775 blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show the up-regulation of the ab4775 phospho signal upon stimulation with GSK-3 beta. Pep
    • Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)
      Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)
      All lanes : Anti-EIF2B5 (phospho S539) antibody (ab4775) at 1/2000 dilution

      Lane 1 : L6 whole cell extract
      Lane 2 : L6 differentiated to myotubes
      Lane 3 : L6 treated with Insulin (10nM for 10 min)
      Lane 4 : C2C12
      Lane 5 : C2C12 treated with Insulin (10nM for 10 min)

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

      Predicted band size: 82 kDa



      A 80 kDa band corresponding to Phospho-eIF2b epsilon was observed across the cell lines tested and reduced upon treatment and in differentiated cell line.

    参考文献 (7)

    ab4775 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。

    ab4775 は 7 報の論文で使用されています。

    • Lim JA  et al. Therapeutic Benefit of Autophagy Modulation in Pompe Disease. Mol Ther 26:1783-1796 (2018). PubMed: 29804932
    • Nichols K  et al. Exogenous essential amino acids stimulate an adaptive unfolded protein response in the mammary glands of lactating cows. J Dairy Sci 100:5909-5921 (2017). PubMed: 28478014
    • Ayuso MI  et al. Stress Granule Induction after Brain Ischemia Is Independent of Eukaryotic Translation Initiation Factor (eIF) 2a Phosphorylation and Is Correlated with a Decrease in eIF4B and eIF4E Proteins. J Biol Chem 291:27252-27264 (2016). PubMed: 27836976
    • Doelman J  et al. Essential amino acid infusions stimulate mammary expression of eukaryotic initiation factor 2Be but milk protein yield is not increased during an imbalance. J Dairy Sci 98:4499-508 (2015). PubMed: 25912861
    • Doelman J  et al. Branched-chain amino acid and lysine deficiencies exert different effects on mammary translational regulation. J Dairy Sci 98:7846-55 (2015). PubMed: 26342977
    • Sidrauski C  et al. Pharmacological brake-release of mRNA translation enhances cognitive memory. Elife 2:e00498 (2013). WB ; Human . PubMed: 23741617
    • Yung HW  et al. Evidence of placental translation inhibition and endoplasmic reticulum stress in the etiology of human intrauterine growth restriction. Am J Pathol 173:451-62 (2008). WB ; Human . PubMed: 18583310

    レビューと Q&A

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    1-3 of 3 Abreviews or Q&A

    Question

    Customer's email is

    Thank you very much for your cooperation.

    Best

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    Answer

    Thank you for your interest and for passing the customer's details.
    DISCOUNT CODE: 42219
    Expiration date: [31/05/2012]
    I am very pleased to hear you would like to accept our offer and test ab4775 in Mouse. This code will give you 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for Mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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    Question

    Thank you for the reply.

    The customer agreed to proceed with the abreview promotionwith ab4775 for WB with mouse samples. Are you interested in this deal in this case? Do you think that there is cross reactivity with mouse?
    I highly appreciate your assistance.
    Looking forward to your reply.
    Thanks again.
    Have a nice day,

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    Answer

    Thank you for your response.
    I can certainly offer a discount code for testing and this case seems to be stronger since there may be some publication regarding the cross-reactivity with mouse (Western blot application).
    Would you be so kind to pass your customer's details so that I can provide the discount as soon as possible?
    I look forward to hearing from you soon.

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    Question

    Further to previous correspondence. thanks a lot for your response and your suggestions. I recommended your suggestion to block with BSA instead of milk powder to reduce background staining but you would expect that the antibody is more specific for Phospho-elF2B epsilon. To answer your questions: the customer used an antibody for elF2B epsilon from Cell Signaling. Cell Signaling offered also a phospho-specific antibody which is now no longer in their sortiment due to a variety of problems. The cells the customer used for WB contain GSK3, also active in non-stimulated cells (see JBC 280, 33006-33014, 2005). elF2B should be phosphorylated under these conditions. Do you offer a positive control the customer could use for the immunoblotting with this antibody? Thanks a lot in advance for further support! Have a nice day and weekend! Best regards

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    Answer

    Thank you for getting back to me. Thank you for confirming that both eIF2B and GSK3 are expressed in these cells. We recommend a positive control of recombinant eIF-2B epsilon treated with GSK 3 beta and lambda phosphatase or CHO-T cells transfected with human insulin receptor (IR) +/- insulin. Unfortunately we do not cell these as lysates or extracts. I hope this information helps. Please do not hesitate to contact me should you require further assistance.

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