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Epigenetics and Nuclear Signaling DNA / RNA Translation Regulation

Anti-eIF4E (phospho S209)抗体(ab4774)

  • Datasheet
  • SDS
Submit a review Q&A (2)References (1)

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Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody (ab4774)
  • 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: ICC/IF, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-eIF4E (phospho S209)抗体
    参阅全部 eIF4E 一抗
  • 描述

    兔多克隆抗体to eIF4E (phospho S209)
  • 宿主

    Rabbit
  • 特异性

    This phosphorylation site specific antibody is selective for eIF-4E containing a phosphate on serine 209.
  • 经测试应用

    适用于: ICC/IF, WBmore details
  • 种属反应性

    与反应: Rat, Human
    预测可用于: Mouse, Rabbit, a wide range of other species
  • 免疫原

    Synthetic peptide corresponding to Human eIF4E (phospho S209).

  • 阳性对照

    • WB: HeLa , HEK293, L6 cell lysate ICC/IF: U-87 MG cells
  • 常规说明

    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

性能

  • 形式

    Liquid
  • 存放说明

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

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

    Immunogen affinity purified
  • 纯化说明

    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.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Regulation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • RNAi
    • Epigenetics and Nuclear Signaling
    • RNAi
    • Eukaryotic Initiation factors (eIF's)

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human eIF4E protein (ab56276)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4774于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF
1/250.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
说明
ICC/IF
1/250.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

靶标

  • 功能

    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.
  • 序列相似性

    Belongs to the eukaryotic initiation factor 4E family.
  • 翻译后修饰

    Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex.
  • Target information above from: UniProt accession P06730 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 1977 Human
    • Entrez Gene: 13684 Mouse
    • Entrez Gene: 117045 Rat
    • Omim: 133440 Human
    • SwissProt: P06730 Human
    • SwissProt: P63073 Mouse
    • SwissProt: P29338 Rabbit
    • SwissProt: P63074 Rat
    • Unigene: 249718 Human
    • Unigene: 3941 Mouse
    • Unigene: 11275 Rat
    see all
  • 别名

    • 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

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody (ab4774)
    Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody (ab4774)

    Immunofluorescence analysis using 70% confluent log phase U-87 MG cells labeled for Phospho-eIF4E pSer209 using ab4774. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ab4774 at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1:2000 for 45 minutes at room temperature (Panel A: green). Nuclei (Panel B: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel C: red) was stained with Rhodamine Phalloidin (1:300). Panel D is a merged image showing cytoplasmic localization. Panel E is a no primary antibody control. The images were captured at 60X magnification.

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

    Lane 1 : HeLa whole cell extracts
    Lane 2 : Serum starved HeLa
    Lane 3 : HeLa Serum Starved for overnight followed by Serum Released
    Lane 4 : HEK-293
    Lane 5 : HEK-293 treated for 30 minutes with 25 µg/mL of Anisomycin
    Lane 6 : L6
    Lane 7 : L6 treated for 10 minutes with 200 ng/mL of Insulin

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 0.4 µg/ml

    Predicted band size: 25 kDa



    Western blot analysis was performed on HeLa, HEK-293 (human cell lines) and L6 (rat cell line) cell lysates with various treatments, blotted for eIF4E (pS209) using ab4774. Two bands ~ 25 and 28 kDa band corresponding to eIF4E (pS209) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate 

  • 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

实验方案

  • Peptide Competition Experiment
  • Western blotting protocol

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (1)

发表研究结果有使用 ab4774?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab4774 被引用在 1 文献中.

  • 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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1-2 of 2 Abreviews or Q&A

Question

What is the concentration of batch 132987?

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Abcam community

Verified customer

Asked on Nov 18 2005

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.

Read More

Abcam Scientific Support

回复于 Nov 22 2005

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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Abcam community

Verified customer

Asked on Aug 16 2004

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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Abcam Scientific Support

回复于 Aug 18 2004

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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