使用敲除细胞株进行验证重组RabMAb

重组Anti-SCD1抗体[EPR21963] (ab236868)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21963] to SCD1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-SCD1抗体[EPR21963]
    参阅全部 SCD1 一抗

  • 描述

    兔单克隆抗体[EPR21963] to SCD1

  • 宿主

    Rabbit

  • 特异性

    ab236868 is recommended for human only in WB.

  • 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: Mouse, Rat, Human

  • 免疫原

    Recombinant fragment within Human SCD1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: O00767

  • 阳性对照

    • WB: HepG2, HEK-293 and SK-MEL-28 whole cell lysate. IHC-P: Human adipose tissue within cardiac muscle tissue; Mouse adipose tissue of stomach tissue; Rat adipose tissue of pancreas tissue. ICC/IF: HepG2 and SK-MEL-28 cells. Flow: HepG2 cells. IP: HepG2 and HEK-293 whole cell lysate.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab236868 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Predicted molecular weight: 42 kDa.

ab236868 is recommended for human only in WB.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/500.
IP 1/30.

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

    see all

  • 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 [EPR21963] (ab236868)
    Western blot - Anti-SCD1 antibody [EPR21963] (ab236868)
    All lanes : Anti-SCD1 antibody [EPR21963] (ab236868) 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.

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

    Predicted band size: 42 kDa



    anes 1-3: Merged signal (red and green). Green - ab236868. Red - loading control ab8245 observed at 50 kDa.

    ab236868 Anti-SCD1 antibody [EPR21963] was shown to specifically react with SCD1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265220 (knockout cell lysate ab257658) was used. Wild-type and SCD1 knockout samples were subjected to SDS-PAGE. ab236868 and Anti-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 10000 dilution for 1 hour at room temperature before imaging.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (ab236868)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling SCD1 with ab236868 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution. Nuclear counterstain is DAPI.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)

    Immunohistochemical analysis of paraffin-embedded human adipose tissue within cardiac muscle tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human adipose cells in cardiac muscle (PMID: 15907797) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Western blot - Anti-SCD1 antibody [EPR21963] (ab236868)
    Western blot - Anti-SCD1 antibody [EPR21963] (ab236868)
    All lanes : Anti-SCD1 antibody [EPR21963] (ab236868) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
    Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
    Lane 3 : SK-MEL-28 (human malignant melanoma), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 42 kDa


    Exposure time: 4 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 20876744; PMID: 9843580; PMID: 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

    We recommend that customers do not boil samples to prevent protein aggregation.

  • Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)

    SCD1 was immunoprecipitated from 0.35 mg of HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate with ab236868 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: HEK-293 whole cell lysate 10 μg (input).
    Lane 2: ab236868 IP in HEK-293 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236868 in HEK-293 whole cell lysate (-).

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 20876744; PMID: 9843580; PMID: 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

  • Flow Cytometry - Anti-SCD1 antibody [EPR21963] (ab236868)
    Flow Cytometry - Anti-SCD1 antibody [EPR21963] (ab236868)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cell line labeling SCD1 with ab236868 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.

  • Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunocytochemistry/ Immunofluorescence - Anti-SCD1 antibody [EPR21963] (ab236868)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 (Human malignant melanoma) cells labeling SCD1 with ab236868 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in SK-MEL-28 cell line. Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution. Nuclear counterstain is DAPI.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)

    Immunohistochemical analysis of paraffin-embedded rat adipose tissue of pancreas tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in adipose cells in rat pancreas (PMID: 11500518) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SCD1 antibody [EPR21963] (ab236868)

    Immunohistochemical analysis of paraffin-embedded mouse adipose tissue of stomach tissue labeling SCD1 with ab236868 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in adipose cells in mouse stomach (PMID: 11500518) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)
    Immunoprecipitation - Anti-SCD1 antibody [EPR21963] (ab236868)

    SCD1 was immunoprecipitated from 10 μg of HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with ab236868 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236868 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: HepG2 whole cell lysate 10 μg (input).
    Lane 2: ab236868 IP in HepG2 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236868 in HepG2 whole cell lysate (-).

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID: 20876744; PMID: 9843580; PMID: 17449569). The full-length protein migrates at 37 kDa; the 28 kDa fragment may represent an SCD1 cleavage product.

  • Anti-SCD1 antibody [EPR21963] (ab236868)
    Anti-SCD1 antibody [EPR21963] (ab236868)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (1)

发表研究结果有使用 ab236868？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab236868 被引用在 1 文献中.

  • Dong Y  et al. (5R)-5-hydroxytriptolide ameliorates liver lipid accumulation by suppressing lipid synthesis and promoting lipid oxidation in mice. Life Sci 232:116644 (2019). PubMed: 31301418

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