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使用敲除细胞株进行验证重组RabMAb

重组Anti-Lipoamide Dehydrogenase抗体[EPR6635] (ab133551)

Reviews (1) Submit a question References (16)
Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6635] to Lipoamide Dehydrogenase
  • Suitable for: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

概述

  • 产品名称

    Anti-Lipoamide Dehydrogenase抗体[EPR6635]
    参阅全部 Lipoamide Dehydrogenase 一抗

  • 描述

    兔单克隆抗体[EPR6635] to Lipoamide Dehydrogenase

  • 宿主

    Rabbit

  • 经测试应用

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

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Jurkat, HeLa, 293T, MCF7, and Caco-2 cell lysates. IHC-P: Human kidney and Human colon tissues

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab133551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
1/200.
WB (1)
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
说明
ICC/IF
1/200.
WB
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
应用说明
Is unsuitable for Flow Cyt or IP.

靶标

  • 功能

    Lipoamide dehydrogenase is a component of the glycine cleavage system as well as of the alpha-ketoacid dehydrogenase complexes. Involved in the hyperactivation of spermatazoa during capacitation and in the spermatazoal acrosome reaction.

  • 疾病相关

    Note=Defects in DLD are involved in the development of congenital infantile lactic acidosis.
    Defects in DLD are a cause of maple syrup urine disease (MSUD) [MIM:248600]. MSUD is characterized by mental and physical retardation, feeding problems and a maple syrup odor to the urine. The keto acids of the branched-chain amino acids are present in the urine, resulting from a block in oxidative decarboxylation.

  • 序列相似性

    Belongs to the class-I pyridine nucleotide-disulfide oxidoreductase family.

  • 翻译后修饰

    Tyrosine phosphorylated.

  • 细胞定位

    Mitochondrion matrix.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • Dehydrogenase complex, E3 component antibody
    • Diaphorase antibody
    • Dihydrolipoamide dehydrogenase antibody
    • Dihydrolipoyl dehydrogenase antibody
    • Dihydrolipoyl dehydrogenase mitochondrial antibody
    • dld antibody
    • DLDD antibody
    • DLDH antibody
    • DLDH_HUMAN antibody
    • E3 antibody
    • E3 branched chain aplha-keto acid antibody
    • E3 component of pyruvate dehydrogenase antibody
    • E3 component of pyruvate dehydrogenase complex 2 oxo glutarate complex branched chain keto acid dehydrogenase complex antibody
    • GCSL antibody
    • Glycine cleavage system L protein antibody
    • Glycine cleavage system protein L antibody
    • LAD antibody
    • lipoamide dehydrogenase antibody
    • Lipoamide reductase antibody
    • Lipoyl dehydrogenase antibody
    • mitochondrial antibody
    • OTTHUMP00000206744 antibody
    • OTTHUMP00000206746 antibody
    • OTTHUMP00000206748 antibody
    • OTTHUMP00000206749 antibody
    • PHE 3 antibody
    • PHE3 antibody
    • Pyruvate dehydrogenase component E3 antibody
    see all

图片

  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : DLD knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 54 kDa
    Observed band size: 56 kDa why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab133551 observed at 56 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab133551 Anti-Lipoamide Dehydrogenase antibody [EPR6635] was shown to specifically react with Lipoamide Dehydrogenase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265535 (knockout cell lysate ab257922) was used. Wild-type and Lipoamide Dehydrogenase knockout samples were subjected to SDS-PAGE. ab133551 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

    Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Lipoamide Dehydrogenase with ab133551 antibody at a dilution of 1/100.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte from peripheral blood) labeling Lipoamide Dehydrogenase with ab133551 at 1/200 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
     
    Confocal image showing Mitochondria staining in Jurkat cell line.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling Lipoamide Dehydrogenase with ab133551 at 1/200 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
     
    Confocal image showing Mitochondria staining in RAW 264.7 cell line.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  • Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) labeling Lipoamide Dehydrogenase with ab133551 at 1/200 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
     
    Confocal image showing Mitochondria staining in RAW 264.7 cell line.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse heart tissue lysate
    Lane 3 : Mouse kidney tissue lysate
    Lane 4 : Mouse spleen tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 54 kDa
    Observed band size: 54 kDa


    Exposure time: 1 second


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/1000 dilution

    Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
    Lane 2 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage)
    Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell)
    Lane 4 : NIH/3T3 (mouse embryonic fibroblast)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 54 kDa
    Observed band size: 54 kDa


    Exposure time: 1 second


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : MCF7 cell lysate
    Lane 5 : Caco-2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 54 kDa
    Observed band size: 56 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

    Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Lipoamide Dehydrogenase with ab133551 antibody at a dilution of 1/100.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
    Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

文献 (16)

发表研究结果有使用 ab133551？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab133551 被引用在 16 文献中.

  • Sheng G  et al. P2RX7 promotes osteosarcoma progression and glucose metabolism by enhancing c-Myc stabilization. J Transl Med 21:132 (2023). PubMed: 36803784
  • Palmieri EM  et al. Pyruvate dehydrogenase operates as an intramolecular nitroxyl generator during macrophage metabolic reprogramming. Nat Commun 14:5114 (2023). PubMed: 37607904
  • Hossain AJ  et al. Association of Phosphorylated Pyruvate Dehydrogenase with Pyruvate Kinase M2 Promotes PKM2 Stability in Response to Insulin. Int J Mol Sci 24:N/A (2023). PubMed: 37761999
  • Beà A  et al. Cardiac fibroblasts display endurance to ischemia, high ROS control and elevated respiration regulated by the JAK2/STAT pathway. FEBS J 289:2540-2561 (2022). PubMed: 34796659
  • Zhang R  et al. An NF-κB/OVOL2 circuit regulates glucose import and cell survival in non-small cell lung cancer. Cell Commun Signal 20:40 (2022). PubMed: 35346238
View all Publications for this product

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Western blot abreview for Anti-Lipoamide Dehydrogenase antibody [EPR6635]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - other (Striatal cell line, pheochromocytoma cell line)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris gradient gel. Laemmli buffer with 10% β-Mercaptoethanol. Samples were denatured at 55C for 10 min. Immobilon-FL transfer membranes. Overnight transfer at +4C.)
Loading amount
20 µg
Specification
Striatal cell line, pheochromocytoma cell line
Blocking step
SEA Block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jan 04 2018

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