Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NSDHL antibody [EPR14489(2)] (ab199730)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14489(2)] to NSDHL
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NSDHL antibody [EPR14489(2)]
    See all NSDHL primary antibodies

  • Description

    Rabbit monoclonal [EPR14489(2)] to NSDHL

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HEK293T, A431, HeLa, HepG2, NIH/3T3, PC-12 and C6 whole cell lysates; Human fetal brain lysate. IP: Human fetal brain whole cell lysate.

  • General notes

     

     

    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.

Properties

Images

  • Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    All lanes : Anti-NSDHL antibody [EPR14489(2)] (ab199730) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : NSDHL knockout HEK293T cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : HeLa 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
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab199730 observed at 38 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab199730 Anti-NSDHL antibody [EPR14489(2)] was shown to specifically react with NSDHL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266682 (knockout cell lysate ab258082) was used. Wild-type and NSDHL knockout samples were subjected to SDS-PAGE. ab199730 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.

  • Immunoprecipitation - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Immunoprecipitation - Anti-NSDHL antibody [EPR14489(2)] (ab199730)

    NSDHL was immunoprecipitated from 1mg of Human fetal brain whole cell lysate with ab199730 at 1/50 dilution.

    Lane 1: Human fetal brain whole cell lysate 10ug (Input).

    Lane 2: ab199730 IP in Human fetal brain whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199730 in Human fetal brain whole cell lysate.

    Western blot was performed from the immunoprecipitate using ab199730 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

  • Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    All lanes : Anti-NSDHL antibody [EPR14489(2)] (ab199730) at 1/1000 dilution

    Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 4 : Human fetal brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Western blot - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    All lanes : Anti-NSDHL antibody [EPR14489(2)] (ab199730) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : C6 (Rat glial tumor cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 10 seconds


    Blocking buffer: 5% NFDM/TBST.

    Dilution buffer: 5% NFDM /TBST or 1%BSA /TBST.

  • Immunoprecipitation - Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Immunoprecipitation - Anti-NSDHL antibody [EPR14489(2)] (ab199730)

    NSDHL was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab199730 at 1/50 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input).

    Lane 2: ab199730 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199730 in HeLa whole cell lysate.

    Western blot was performed from the immunoprecipitate using ab199730 at 1/1000 dilution (Panel A) or ab190353 at 1/1000 dilution (Panel B).

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds (Panel A and B).

     ab199730 was used to immunoprecipitate NSDHL. These precipitates were resolved with ab190353.

  • Anti-NSDHL antibody [EPR14489(2)] (ab199730)
    Anti-NSDHL antibody [EPR14489(2)] (ab199730)

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