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遺伝子ノックアウト細胞株 検証済

Anti-MTHFD2 抗体 [4G7-2G3] (ab56772)

Submit a review Q&A (1)References (10)
Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
  • Flow Cytometry - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
  • Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)

Key features and details

  • Mouse monoclonal [4G7-2G3] to MTHFD2
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2a

製品の概要

  • 製品名

    Anti-MTHFD2 antibody [4G7-2G3]
    MTHFD2 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [4G7-2G3] to MTHFD2

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cytmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant full length protein, corresponding to amino acids 16-345 of Human MTHFD2

  • ポジティブ･コントロール

    • WB: HEK293T, HepG2 and HeLa cell lysates. IHC-P: Human lymphoma tissue. Flow Cyt: HepG2 cells.

  • 特記事項

    This product was changed from ascites to tissue culture supernatant on 24/1/19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    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ab56772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
Use at an assay dependent concentration. Predicted molecular weight: 37 kDa.
IHC-P
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. 
特記事項
WB
Use at an assay dependent concentration. Predicted molecular weight: 37 kDa.
IHC-P
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. 

ターゲット情報

  • 配列類似性

    Belongs to the tetrahydrofolate dehydrogenase/cyclohydrolase family.

  • 発生段階

    Expressed only in developing normal tissues.

  • 細胞内局在

    Mitochondrion.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial antibody
    • Descriptions antibody
    • Methenyltetrahydrofolate cyclohydrolase antibody
    • methylene tetrahydrofolate dehydrogenase (NAD+ dependent) antibody
    • methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2 antibody
    • methylenetetrahydrofolate dehydrogenase (NADP+ dependent) antibody
    • methylenetetrahydrofolate dehydrogenase 2 antibody
    • MGC82516 antibody
    • MTDC_HUMAN antibody
    • MTHFD2 antibody
    • mthfd2 methylene tetrahydrofolate dehydrogenase (NAD+ dependent) antibody
    • NAD-dependent methylene tetrahydrofolate dehydrogenase antibody
    • NAD-dependent methylenetetrahydrofolate dehydrogenase antibody
    • NMDMC antibody
    see all

画像

  • Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
    Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
    All lanes : Anti-MTHFD2 antibody [4G7-2G3] (ab56772) at 1/500 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : MTHFD2 knockout HEK293T cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

    Predicted band size: 37 kDa
    Observed band size: 36 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab56772 observed at 36 kDa. Red - loading control ab52901 observed at  kDa.

     ab56772 Anti-MTHFD2 antibody was shown to specifically react with MTHFD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266383 (knockout cell lysate ab258528) was used. Wild-type and MTHFD2 knockout samples were subjected to SDS-PAGE. ab56772 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)

    MTHFD2 antibody (ab56772) used in immunohistochemistry at 3μg/ml on formalin fixed and paraffin embedded human lymphoma tissue.

  • Flow Cytometry - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
    Flow Cytometry - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)

    Overlay histogram showing HepG2 cells stained with ab56772 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56772, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    This image was generated using the ascites version of the product.

  • Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)
    Western blot - Anti-MTHFD2 antibody [4G7-2G3] (ab56772)

    MTHFD2 antibody (ab56772) at 1μg/lane + HepG2 cell lysate at 25μg/lane.

    This image was generated using the ascites version of the product.

参考文献 (10)

ab56772 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。

ab56772 は 10 報の論文で使用されています。

  • Green AC  et al. Formate overflow drives toxic folate trapping in MTHFD1 inhibited cancer cells. Nat Metab 5:642-659 (2023). PubMed: 37012496
  • Tanaka K  et al. Glioma cells require one-carbon metabolism to survive glutamine starvation. Acta Neuropathol Commun 9:16 (2021). PubMed: 33468252
  • Nicolaidou V  et al. Detection and characterisation of novel alternative splicing variants of the mitochondrial folate enzyme MTHFD2. Mol Biol Rep 47:7089-7096 (2020). PubMed: 32880830
  • Nishimura T  et al. Cancer stem-like properties and gefitinib resistance are dependent on purine synthetic metabolism mediated by the mitochondrial enzyme MTHFD2. Oncogene 38:2464-2481 (2019). PubMed: 30532069
  • Green NH  et al. MTHFD2 links RNA methylation to metabolic reprogramming in renal cell carcinoma. Oncogene 38:6211-6225 (2019). PubMed: 31289360
  • Gustafsson R  et al. Crystal Structure of the Emerging Cancer Target MTHFD2 in Complex with a Substrate-Based Inhibitor. Cancer Res 77:937-948 (2017). PubMed: 27899380
  • Pikman Y  et al. Targeting MTHFD2 in acute myeloid leukemia. J Exp Med 213:1285-306 (2016). PubMed: 27325891
  • Koufaris C  et al. Systematic integration of molecular profiles identifies miR-22 as a regulator of lipid and folate metabolism in breast cancer cells. Oncogene 35:2766-76 (2016). PubMed: 26477310
  • Gustafsson Sheppard N  et al. The folate-coupled enzyme MTHFD2 is a nuclear protein and promotes cell proliferation. Sci Rep 5:15029 (2015). WB . PubMed: 26461067
  • Nilsson R  et al. Metabolic enzyme expression highlights a key role for MTHFD2 and the mitochondrial folate pathway in cancer. Nat Commun 5:3128 (2014). WB ; Human . PubMed: 24451681

レビューと Q&A

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Question

will test ab56772 in flow cytometry

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Answer

Thank you for your phone call.


DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab56772 in flow cytometry. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for flow cytometry 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: https://www.abcam.com/collaborationdiscount.

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