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使用敲除细胞株进行验证

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

概述

  • 产品名称

    Anti-MTHFD2抗体[4G7-2G3]
    参阅全部 MTHFD2 一抗

  • 描述

    小鼠单克隆抗体[4G7-2G3] to MTHFD2

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, IHC-P, Flow Cytmore details

  • 种属反应性

    与反应: Human

  • 免疫原

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

  • 阳性对照

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

  • 常规说明

    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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab56772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
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. 
说明
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. 

靶标

  • 序列相似性

    Belongs to the tetrahydrofolate dehydrogenase/cyclohydrolase family.

  • 发展阶段

    Expressed only in developing normal tissues.

  • 细胞定位

    Mitochondrion.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • 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.

文献 (10)

发表研究结果有使用 ab56772？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab56772 被引用在 10 文献中.

  • 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

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will test ab56772 in flow cytometry

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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.

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