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使用敲除细胞株进行验证

Anti-ETFA抗体[2B11AE8] (ab110316)

Submit a review Submit a question References (8)
Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
  • Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
  • Immunocytochemistry/ Immunofluorescence - Anti-ETFA antibody [2B11AE8] (ab110316)
  • Flow Cytometry - Anti-ETFA antibody [2B11AE8] (ab110316)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETFA antibody [2B11AE8] (ab110316)

Key features and details

  • Mouse monoclonal [2B11AE8] to ETFA
  • Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG2b

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647

概述

  • 产品名称

    Anti-ETFA抗体[2B11AE8]
    参阅全部 ETFA 一抗

  • 描述

    小鼠单克隆抗体[2B11AE8] to ETFA

  • 宿主

    Mouse

  • 经测试应用

    适用于: WB, ICC/IF, Flow Cyt, IHC-Pmore details

  • 种属反应性

    与反应: Mouse, Rat, Cow, Human

  • 免疫原

    Recombinant full length protein. This information is considered to be commercially sensitive.

  • 阳性对照

    • ICC: HeLa cells. Flow Cyt: HL-60 cells. IHC-P: Human heart tissue. WB: HepG2 and HEK-293T whole cell lysates; Human liver mitochondria; Bovine heart mitochondria; Rat liver mitochondria; Mouse liver mitochondria.

  • 常规说明

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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

    Product was previously marketed under the MitoSciences sub-brand.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab110316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 a concentration of 1 µg/ml. Predicted molecular weight: 35 kDa.
ICC/IF
Use a concentration of 4 µg/ml.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 5 µg/ml.
说明
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 35 kDa.
ICC/IF
Use a concentration of 4 µg/ml.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 5 µg/ml.

靶标

  • 功能

    The electron transfer flavoprotein serves as a specific electron acceptor for several dehydrogenases, including five acyl-CoA dehydrogenases, glutaryl-CoA and sarcosine dehydrogenase. It transfers the electrons to the main mitochondrial respiratory chain via ETF-ubiquinone oxidoreductase (ETF dehydrogenase).

  • 疾病相关

    Glutaric aciduria 2A

  • 序列相似性

    Belongs to the ETF alpha-subunit/FixB family.

  • 结构域

    Domain I shares an identical polypeptide fold with the beta subunit ETFB though there is no sequence similarity.

  • 翻译后修饰

    The N-terminus is blocked.

  • 细胞定位

    Mitochondrion matrix.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • Alpha ETF antibody
    • Alpha-ETF antibody
    • Electron transfer flavoprotein alpha polypeptide antibody
    • Electron transfer flavoprotein alpha subunit antibody
    • Electron transfer flavoprotein subunit alpha mitochondrial antibody
    • Electron transfer flavoprotein subunit alpha, mitochondrial antibody
    • Electron transferring flavoprotein alpha polypeptide antibody
    • EMA antibody
    • ETFA antibody
    • ETFA_HUMAN antibody
    • GA2 antibody
    • Glutaric aciduria II antibody
    see all

图片

  • Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
    Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
    All lanes : Anti-ETFA antibody [2B11AE8] (ab110316) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : ETFA knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate

    Lysates/proteins at 40 µg per lane.

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

    Predicted band size: 35 kDa
    Observed band size: 35 kDa



    Lanes 1-2: Merged signal (red and green). Green - ab110316 observed at 35 kDa. Red - loading control ab52901 observed at  kDa.

     ab110316 Anti-ETFA antibody [2B11AE8]  was shown to specifically react with ETFA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266513 (knockout cell lysate ab257943) was used. Wild-type and ETFA knockout samples were subjected to SDS-PAGE. ab110316 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 1000 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.

  • Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
    Western blot - Anti-ETFA antibody [2B11AE8] (ab110316)
    All lanes : Anti-ETFA antibody [2B11AE8] (ab110316) at 1 µg/ml

    Lane 1 : Human heart tissue
    Lane 2 : HepG2 whole cells
    Lane 3 : Human liver mitochondria
    Lane 4 : Bovine heart mitochondria
    Lane 5 : Rat liver mitochondria
    Lane 6 : Mouse liver mitochondria

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 35 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ETFA antibody [2B11AE8] (ab110316)
    Immunocytochemistry/ Immunofluorescence - Anti-ETFA antibody [2B11AE8] (ab110316)
    Immunocytochemistry image of stained HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the antibody (ab110316, 4 µg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. The target protein locates to the mitochondrial matrix
  • Flow Cytometry - Anti-ETFA antibody [2B11AE8] (ab110316)
    Flow Cytometry - Anti-ETFA antibody [2B11AE8] (ab110316)
    HL-60 cells were stained with 1 µg/mL ab110316 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETFA antibody [2B11AE8] (ab110316)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETFA antibody [2B11AE8] (ab110316)

    IHC image of ETFA staining in Human heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110316, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

文献 (8)

发表研究结果有使用 ab110316？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab110316 被引用在 8 文献中.

  • Andonian BJ  et al. Rheumatoid arthritis T cell and muscle oxidative metabolism associate with exercise-induced changes in cardiorespiratory fitness. Sci Rep 12:7450 (2022). PubMed: 35523821
  • Fiorentino TV  et al. Pioglitazone corrects dysregulation of skeletal muscle mitochondrial proteins involved in ATP synthesis in type 2 diabetes. Metabolism 114:154416 (2021). PubMed: 33137378
  • Ehrlicher SE  et al. Mitochondrial adaptations to exercise do not require Bcl2-mediated autophagy but occur with BNIP3/Parkin activation. FASEB J 34:4602-4618 (2020). PubMed: 32030805
  • Fisher-Wellman KH  et al. Respiratory Phenomics across Multiple Models of Protein Hyperacylation in Cardiac Mitochondria Reveals a Marginal Impact on Bioenergetics. Cell Rep 26:1557-1572.e8 (2019). PubMed: 30726738
  • White PJ  et al. The BCKDH Kinase and Phosphatase Integrate BCAA and Lipid Metabolism via Regulation of ATP-Citrate Lyase. Cell Metab 27:1281-1293.e7 (2018). PubMed: 29779826
  • Stauch KL  et al. Loss of Pink1 modulates synaptic mitochondrial bioenergetics in the rat striatum prior to motor symptoms: concomitant complex I respiratory defects and increased complex II-mediated respiration. Proteomics Clin Appl 10:1205-1217 (2016). PubMed: 27568932
  • Davoudi M  et al. Complex I function and supercomplex formation are preserved in liver mitochondria despite progressive complex III deficiency. PLoS One 9:e86767 (2014). WB ; Mouse . PubMed: 24466228
  • Pougovkina O  et al. Mitochondrial protein acetylation is driven by acetyl-CoA from fatty acid oxidation. Hum Mol Genet N/A:N/A (2014). Mouse . PubMed: 24516071

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