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Anti-ECSIT 抗体 (ab21288)

Submit a review Q&A (3)References (7)
Western blot - Anti-ECSIT antibody (ab21288)

    Key features and details

    • Rabbit polyclonal to ECSIT
    • Suitable for: IHC-P, IP, ICC/IF, WB
    • Reacts with: Mouse, Human
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-ECSIT antibody

    • 製品の詳細

      Rabbit polyclonal to ECSIT

    • 由来種

      Rabbit

    • アプリケーション

      適用あり: IHC-P, IP, ICC/IF, WBmore details

    • 種交差性

      交差種: Mouse, Human

    • 免疫原

      Synthetic peptide corresponding to 14 amino acids near the C-terminus of ECSIT (Human).

    • 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

    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ab21288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
    IHC-P
    Use at an assay dependent concentration.
    IP
    Use at an assay dependent concentration. PubMed: 21163940
    ICC/IF
    Use at an assay dependent concentration. PubMed: 21163940
    WB
    Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 49 kDa).
    特記事項
    IHC-P
    Use at an assay dependent concentration.
    IP
    Use at an assay dependent concentration. PubMed: 21163940
    ICC/IF
    Use at an assay dependent concentration. PubMed: 21163940
    WB
    Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 49 kDa).

    ターゲット情報

    • 機能

      Adapter protein of the Toll-like and IL-1 receptor signaling pathway that is involved in the activation of NF-kappa-B via MAP3K1. Promotes proteolytic activation of MAP3K1. Involved in the BMP signaling pathway. Required for normal embryonic development (By similarity). Required for efficient assembly of mitochondrial NADH:ubiquinone oxidoreductase.

    • 配列類似性

      Belongs to the ECSIT family.

    • 細胞内局在

      Cytoplasm. Nucleus. Mitochondrion.
    • Information by UniProt

    • 参照データベース

      • 別名

        • Ecsit antibody
        • ECSIT homolog antibody
        • ECSIT protein antibody
        • ECSIT_HUMAN antibody
        • Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial antibody
        • Evolutionarily conserved signaling intermediate in Toll pathways antibody
        • Likely ortholog of mouse signaling intermediate in Toll pathway evolutionarily conserved antibody
        • Protein SITPEC antibody
        • Signaling intermediate in Toll pathway evolutionarily conserved antibody
        • SITPEC antibody
        • SITPEC protein antibody
        see all

      画像

      • Western blot - Anti-ECSIT antibody (ab21288)
        Western blot - Anti-ECSIT antibody (ab21288)
        Lane 1 : Anti-ECSIT antibody (ab21288) at 0.5 µg/ml
        Lane 2 : Anti-ECSIT antibody (ab21288) at 1 µg/ml
        Lane 3 : Anti-ECSIT antibody (ab21288) at 2 µg/ml

        All lanes : human heart lysate

        Predicted band size: 49 kDa
        Observed band size: 42 kDa why is the actual band size different from the predicted?

      参考文献 (7)

      ab21288 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。

      ab21288 は 7 報の論文で使用されています。

      • Marrocco A  et al. Metabolic Adaptation of Macrophages as Mechanism of Defense against Crystalline Silica. J Immunol 207:1627-1640 (2021). PubMed: 34433619
      • Meng H  et al. Synthetic VSMCs induce BBB disruption mediated by MYPT1 in ischemic stroke. iScience 24:103047 (2021). PubMed: 34553133
      • Seo JH  et al. MTFMT deficiency correlates with reduced mitochondrial integrity and enhanced host susceptibility to intracellular infection. Sci Rep 10:11183 (2020). PubMed: 32636430
      • Kim MJ  et al. CRBN Is a Negative Regulator of Bactericidal Activity and Autophagy Activation Through Inhibiting the Ubiquitination of ECSIT and BECN1. Front Immunol 10:2203 (2019). PubMed: 31620128
      • Guarani V  et al. TIMMDC1/C3orf1 functions as a membrane-embedded mitochondrial complex I assembly factor through association with the MCIA complex. Mol Cell Biol 34:847-61 (2014). PubMed: 24344204
      • Soler-López M  et al. Interactome mapping suggests new mechanistic details underlying Alzheimer's disease. Genome Res 21:364-76 (2011). WB, ICC/IF, IP ; Human . PubMed: 21163940
      • Vogel RO  et al. Cytosolic signaling protein Ecsit also localizes to mitochondria where it interacts with chaperone NDUFAF1 and functions in complex I assembly. Genes Dev 21:615-24 (2007). PubMed: 17344420

      レビューと Q&A

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      レビューを送る 質問を送る

      1-3 of 3 Abreviews or Q&A

      Question

      Dear Provider,

      we are extensively using your Anti-ECSIT antibody (ab21288), which is giving us high quality results. Nevertheless, we are also experiencing some intriguing results, which may account for post-translational modifications that ECSIT might undergo. Unfortunately, since the sequence of the antibody is not provided (just that is 14 amino acids near the C-terminus), we cannot prove our hypotheses. We would be grateful if you can provide us this information in order to complete our experiments.

      We thank you very much in advance.
      Best,

      Read More
      Answer

      Thank you for your enquiry and your interest.

      I regret to inform you that the immunogen sequence is commercially sensitive information and we are not allowed to release it. However, it is between residues 301 and 350 aa accession number NP_057665.

      Since antibody is a polyclonal antibody, it recognizes different epitopes so epitope mapping is not performed.

      If you need any further assistance, please do not hesitate to contact me.

      Read More

      Question

      Please see reply from Nezira below. regards --- Hi Michelle, i have just received your email about the antibody from abcam, I am sorry but I dont have the lot number from the previous vial as i bought it about 1 year ago and have already used it all. The samples that i tested it on would have been the same because i am still using the same plasmids so the myc tag would have not affected it. I would be happy to take the replacement as they are still the only supplier for this antibody and i am eager to continue with the experiments.I would also be happy to receive any help on how to improve the signal because even with the previous vial that i bought i did not get the same band as they have on the data sheet which comes with the antibody.I am mainly working on the HEK 293 cells but am also looking at PBMCs and CD14positive and negative cells. Thanks Michelle for your help.

      Read More
      Answer

      Thank you very much for your reply and Nezira's. At this stage I think it would be worth looking at Nezira's protocol in detail in light of the fact that the first lot did not give her the results expected. Can you please ask Nezira to click on the link below and fill in our online questionnaire for WB, we will then be able to review all her data and understand the source of the problem. I would already recommend to run a positive control of human heart lysate to make sure that the problem is not due to her sample preparation. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=21288&mode=questionaire We look forward to receiving this information to resolve this matter, thank you both in advance,

      Read More

      Question

      BATCH NUMBER 130274 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Wrong band. It detects a band that looks like it is the correct size, but when truncation mutants of ECSIT were made the band does not change size. SAMPLE Recombinant protein PRIMARY ANTIBODY Abcam ab21288, Diluted 1:1000 in TBST containing 2% BSA 0.01% NaN3. Incubated overnight at 4 degrees C. Followed by 3 quick washes, 3 15-min washes and 3 quick washes. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Negative: Recombinant GFP and untransfected cells. Positive: Recombinant tagged ECSIT (of various lengths due to ECSIT truncation mutatants) ANTIBODY STORAGE CONDITIONS 4 Degrees C, as it arrived SAMPLE PREPARATION SDS Loading Buffer, 3 min at 95 degrees C AMOUNT OF PROTEIN LOADED 1 well of 24-well plate expressing protein. This is what I generally use for western blotting and works well for other combinations of recombinant proteins and respective antibodies. ELECTROPHORESIS/GEL CONDITIONS SDS PAGE, 4-12% Novex Bis Tris TRANSFER AND BLOCKING CONDITIONS For transfer: Invitrogen NuPage Transfer For Blocking: TBS-Tween (0.15M NaCl, 0.1 M Tris HCl pH7.4, 0.1% Tween 20) with 3% milk SECONDARY ANTIBODY [a competitor] goat anti-rabbit HRP conjugated 1:5000 Incubated 2 hours at 25 degrees C. Followed by 3 quick washes, 3 15-min washes and 3 quick washes. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Different cell lysates from independent transfections. Used fresh primary and secondary antibody dilutions every time. ADDITIONAL NOTES Uncertain which of the two is the order number: S102487 or 4500103431

      Read More
      Answer

      Thank you again for your enquiry. I spoke to a colleague of mine who also agreed that you may be detecting the endogenous protein. Did you cleave the tags? The tags may be adding the additional kDa’s to your bands. Another possibility is that differences in the size could be negligible enough that they do not register on your gel. There just is not enough information on your truncation methods to determine where the problem lies. Where is your truncated sample cleaved? The immunogen is within residues 301-350and I hope this information can help you. Please do not hesitate to contact us if you need any more advice or information.

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