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使用敲除细胞株进行验证

Anti-SCD1抗体[CD.E10] (ab19862)

Reviews (6)Q&A (11)References (89)
Western blot - Anti-SCD1 antibody [CD.E10] (ab19862)
  • Immunoprecipitation - Anti-SCD1 antibody [CD.E10] (ab19862)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [CD.E10] (ab19862)
  • Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [CD.E10] (ab19862)
  • Flow Cytometry - Anti-SCD1 antibody [CD.E10] (ab19862)

Key features and details

  • Mouse monoclonal [CD.E10] to SCD1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2b

概述

  • 产品名称

    Anti-SCD1抗体[CD.E10]
    参阅全部 SCD1 一抗

  • 描述

    小鼠单克隆抗体[CD.E10] to SCD1

  • 宿主

    Mouse

  • 特异性

    This SCD1 antibody can bind to SCD5 very weakly in WB when SCD5 is overexpressed (<10% cross-reactivity).

  • 经测试应用

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

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant full length protein (Human).

  • 阳性对照

    • WB: HepG2 and HeLa cell lysates. Flow Cyt: HepG2 cells. IP: Hek293 whole cell extract. IHC: Human skin tissue.

  • 常规说明

    SCD1 is known to undergo post-translational modifications and the sizes differ in different cell lines so the observed band size can be different than predicted band size. As positive control we recommend using SCD1 over-expressed 293 transfected cell lysates for western blot.

    This product was changed from ascites to tissue culture supernatant on 25th May 2018. 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ab19862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 (3)
1/1000. Predicted molecular weight: 42 kDa.
IHC-P (1)
Use a concentration of 4 µg/ml.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (2)
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. 
IP
Use at an assay dependent concentration.
说明
WB
1/1000. Predicted molecular weight: 42 kDa.
IHC-P
Use a concentration of 4 µg/ml.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. 
IP
Use at an assay dependent concentration.

靶标

  • 功能

    Terminal component of the liver microsomal stearyl-CoA desaturase system, that utilizes O(2) and electrons from reduced cytochrome b5 to catalyze the insertion of a double bond into a spectrum of fatty acyl-CoA substrates including palmitoyl-CoA and stearoyl-CoA.

  • 序列相似性

    Belongs to the fatty acid desaturase family.

  • 结构域

    The histidine box domains may contain the active site and/or be involved in metal ion binding.

  • 细胞定位

    Endoplasmic reticulum membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

      • ACOD_HUMAN antibody
      • Acyl-CoA desaturase antibody
      • Delta(9)-desaturase antibody
      • Delta-9 desaturase antibody
      • Delta-9-Desaturase antibody
      • FADS5 antibody
      • Fatty acid desaturase antibody
      • PRO0998 antibody
      • SCD 1 antibody
      • SCD antibody
      • SCD1 antibody
      • Stearoyl-CoA desaturase antibody
      see all

    图片

    • Western blot - Anti-SCD1 antibody [CD.E10] (ab19862)
      Western blot - Anti-SCD1 antibody [CD.E10] (ab19862)
      All lanes : Anti-SCD1 antibody [CD.E10] (ab19862) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : SCD knockout HeLa cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 42 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 - ab19862 observed at 36 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

      ab19862 was shown to react with SCD1 in wild-type HeLa cells in western blot with loss of signal observed in SCD knockout cell line ab265220 (SCD knockout cell lysate ab257658). Wild-type and SCD knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab19862 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunoprecipitation - Anti-SCD1 antibody [CD.E10] (ab19862)
      Immunoprecipitation - Anti-SCD1 antibody [CD.E10] (ab19862)
      SCD was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Mouse monoclonal to SCD and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab19862.
      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
      Band: 32kDa; SCD.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [CD.E10] (ab19862)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [CD.E10] (ab19862)
      ab19862 staining SCD in human skin.
      Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
      Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
    • Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [CD.E10] (ab19862)
      Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [CD.E10] (ab19862)
      ICC/IF image of ab19862 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19862, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Flow Cytometry - Anti-SCD1 antibody [CD.E10] (ab19862)
      Flow Cytometry - Anti-SCD1 antibody [CD.E10] (ab19862)
      Overlay histogram showing HepG2 cells stained with ab19862 (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 (ab19862, 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 IgG2b [PLPV219] (ab91366, 2µg/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 in 0.1% PBS-Tween used under the same conditions.

    文献 (89)

    发表研究结果有使用 ab19862？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab19862 被引用在 89 文献中.

    • Li H  et al. Geniposide plus chlorogenic acid reverses non-alcoholic steatohepatitis via regulation of gut microbiota and bile acid signaling in a mouse model in vivo. Front Pharmacol 14:1148737 (2023). PubMed: 37077819
    • Tovar R  et al. Administration of Linoleoylethanolamide Reduced Weight Gain, Dyslipidemia, and Inflammation Associated with High-Fat-Diet-Induced Obesity. Nutrients 15:N/A (2023). PubMed: 37892524
    • Liu M  et al. Dominant-negative HNF1α mutant promotes liver steatosis and inflammation by regulating hepatic complement factor D. iScience 26:108018 (2023). PubMed: 37841581
    • Wong TL  et al. ADAR1-mediated RNA editing of SCD1 drives drug resistance and self-renewal in gastric cancer. Nat Commun 14:2861 (2023). PubMed: 37208334
    • Bedoya-Guzmán FA  et al. BACE1 and SCD1 are associated with neurodegeneration. Front Aging Neurosci 15:1194203 (2023). PubMed: 37744400
    View all Publications for this product

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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SCD1 antibody [CD.E10]

     Poor
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Skin sections)
    Specification
    Skin sections
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3%
    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 15 2007

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