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使用敲除细胞株进行验证重组RabMAb

重组Anti-GALT抗体[EPR12555] (ab178406)

  • Datasheet
  • SDS
Submit a review Submit a question References (6)

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Western blot - Anti-GALT antibody [EPR12555] (ab178406)
  • Western blot - Anti-GALT antibody [EPR12555] (ab178406)
  • Western blot - Anti-GALT antibody [EPR12555] (ab178406)
  • Immunoprecipitation - Anti-GALT antibody [EPR12555] (ab178406)
  • Anti-GALT antibody [EPR12555] (ab178406)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12555] to GALT
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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概述

  • 产品名称

    Anti-GALT抗体[EPR12555]
    参阅全部 GALT 一抗
  • 描述

    兔单克隆抗体[EPR12555] to GALT
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, IPmore details
  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Rat
  • 免疫原

    Synthetic peptide within Human GALT aa 50-150. The exact sequence is proprietary.
    Database link: P07902

  • 阳性对照

    • WB: HeLa, A549, K562, MDA-MB-435 and HepG2 whole cell lysate (ab7900); Human fetal brain and fetal liver lysates.
  • 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

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EPR12555
  • 同种型

    IgG
  • 研究领域

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism

相关产品

  • Alternative Versions

    • Anti-GALT antibody [EPR12555] - BSA and Azide free (ab240206)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Hep G2 whole cell lysate (ab166833)
  • Recombinant Protein

    • Recombinant Human GALT protein (ab139613)
  • Related Products

    • Recombinant Human GALT protein (ab139613)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab178406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
1/1000 - 1/5000. Predicted molecular weight: 43 kDa.
IP
1/10 - 1/100.
说明
WB
1/1000 - 1/5000. Predicted molecular weight: 43 kDa.
IP
1/10 - 1/100.

靶标

  • 通路

    Carbohydrate metabolism; galactose metabolism.
  • 疾病相关

    Defects in GALT are the cause of galactosemia (GALCT) [MIM:230400]. Galactosemia is an inherited disorder of galactose metabolism that causes jaundice, cataracts, and mental retardation.
  • 序列相似性

    Belongs to the galactose-1-phosphate uridylyltransferase type 1 family.
  • Target information above from: UniProt accession P07902 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 2592 Human
    • Entrez Gene: 14430 Mouse
    • Entrez Gene: 298003 Rat
    • Omim: 606999 Human
    • SwissProt: P07902 Human
    • SwissProt: Q03249 Mouse
    • SwissProt: P43424 Rat
    • Unigene: 522090 Human
    • Unigene: 439669 Mouse
    • Unigene: 102781 Rat
    see all
  • 别名

    • Gal 1 P uridylyltransferase antibody
    • Gal-1-P uridylyltransferase antibody
    • Galactose 1 phosphate uridyl transferase antibody
    • Galactose 1 phosphate uridylyltransferase antibody
    • Galactose-1-phosphate uridylyltransferase antibody
    • GALT antibody
    • GALT_HUMAN antibody
    • UDP glucose hexose 1 phosphate uridylyltransferase antibody
    • UDP-glucose--hexose-1-phosphate uridylyltransferase antibody
    see all

图片

  • Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    All lanes : Anti-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : GALT knockout HeLa cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 43 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab178406 observed at 43 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab178406 Anti-GALT antibody [EPR12555] was shown to specifically react with Galactose-1-Phosphate Uridylyltransferase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265027 (knockout cell lysate ab257958) was used. Wild-type and Galactose-1-Phosphate Uridylyltransferase knockout samples were subjected to SDS-PAGE. ab178406 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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.

     

  • Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    Lanes 1-2 : Anti-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution
    Lanes 3-4 : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 1/2000 dilution

    Lanes 1 & 3 : GALT knockout HAP1 cell lysate
    Lanes 2 & 4 : Wild-type HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 43 kDa



     

    Lanes 1 and 2: Green signal from target – ab178406 observed at 43 kDa

    Lanes 3 and 4: Red signal from loading control – ab8245 observed at 37 kDa

    Lanes 5 and 6: Merged (red and green) signal

    ab178406 was shown to react with GALT when GALT knockout samples were used, along with additional cross-reactive bands. Wild-type and GALT knockout samples were subjected to SDS-PAGE. ab178406 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW)preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    Western blot - Anti-GALT antibody [EPR12555] (ab178406)
    All lanes : Anti-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : MDA-MB-435 cell lysate
    Lane 3 : Human fetal brain lysate
    Lane 4 : Human fetal liver lysate
    Lane 5 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 43 kDa

  • Immunoprecipitation - Anti-GALT antibody [EPR12555] (ab178406)
    Immunoprecipitation - Anti-GALT antibody [EPR12555] (ab178406)

    Western blot analysis on Immunoprecipitation pellet from either 1) Human fetal liver lysate, or 2) 1xPBS (negative control); showing GALT, immunoprecipitated using ab178406 at 1/10 dilution with HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

  • Anti-GALT antibody [EPR12555] (ab178406)
    Anti-GALT antibody [EPR12555] (ab178406)

实验方案

  • Western blot protocols
  • Immunoprecipitation protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (6)

发表研究结果有使用 ab178406?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab178406 被引用在 6 文献中.

  • Delnoy B  et al. Novel mRNA therapy restores GALT protein and enzyme activity in a zebrafish model of classic galactosemia. J Inherit Metab Dis 45:748-758 (2022). PubMed: 35527402
  • Balakrishnan B  et al. Novel mRNA-Based Therapy Reduces Toxic Galactose Metabolites and Overcomes Galactose Sensitivity in a Mouse Model of Classic Galactosemia. Mol Ther 28:304-312 (2020). PubMed: 31604675
  • Tran H  et al. Mast Cells Induce Blood Brain Barrier Damage in SCD by Causing Endoplasmic Reticulum Stress in the Endothelium. Front Cell Neurosci 13:56 (2019). PubMed: 30837844
  • Sun YZ  et al. Proteomic and bioinformatic analysis of condyloma acuminata: mild hyperthermia treatment reveals compromised HPV infectivity of keratinocytes via regulation of metabolism, differentiation and anti-viral responses. Int J Hyperthermia 36:383-393 (2019). PubMed: 30909744
  • Deng Y  et al. Activity of the SPCA1 Calcium Pump Couples Sphingomyelin Synthesis to Sorting of Secretory Proteins in the Trans-Golgi Network. Dev Cell 47:464-478.e8 (2018). PubMed: 30393074
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371

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