Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-NSDHL antibody [EPR14490] (ab190353)

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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 [EPR14490] to NSDHL
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-NSDHL antibody [EPR14490]
    See all NSDHL primary antibodies

  • Description

    Rabbit monoclonal [EPR14490] to NSDHL

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human NSDHL aa 150-250. The exact sequence is proprietary.
    Database link: Q15738

  • Positive control

    • WB: HEK293T, A431, HeLa and Human fetal brain lysates. IHC-P: Human liver and Human adenocarcinoma of endometrium tissue. ICC: U87-MG cells.

  • 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

Applications

Our Abpromise guarantee covers the use of ab190353 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 42 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.

Target

  • Tissue specificity

    Brain, heart, liver, lung, kidney, skin and placenta.

  • Pathway

    Steroid biosynthesis; zymosterol biosynthesis; zymosterol from lanosterol: step 4/6.

  • Involvement in disease

    Defects in NSDHL are the cause of congenital hemidysplasia with ichthyosiform erythroderma and limb defects (CHILD) [MIM:308050]. CHILD is an X-linked dominant disorder of lipid metabolism with disturbed cholesterol biosynthesis, which typically results in male lethality. Clinically, it is characterized by congenital, unilateral, ichthyosisform erythroderma with striking lateralization, sharp midline demarcation, and ipsilateral limb defects and hypoplasia of the body. Limbs defects range from hypoplasia of digits or ribs to complete amelia, often including scoliosis.
    Defects in NSDHL are the cause of CK syndrome (CKS) [MIM:300831]. CKS is a disorder characterized by mild to severe cognitive impairment, seizures, microcephaly, cerebral cortical malformations, dysmorphic facial features, and thin body habitus.

  • Sequence similarities

    Belongs to the 3-beta-HSD family.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • decarboxylating antibody
      • H105E3 antibody
      • H105e3 protein antibody
      • NAD(P) dependent steroid dehydrogenase like antibody
      • NSDHL antibody
      • NSDHL_HUMAN antibody
      • Protein H105e3 antibody
      • SDR31E1 antibody
      • Short chain dehydrogenase/reductase family 31E member 1 antibody
      • Sterol 4 alpha carboxylate 3 dehydrogenase decarboxylating antibody
      • Sterol-4-alpha-carboxylate 3-dehydrogenase antibody
      • XAP104 antibody
      see all

    Images

    • Western blot - Anti-NSDHL antibody [EPR14490] (ab190353)
      Western blot - Anti-NSDHL antibody [EPR14490] (ab190353)
      All lanes : Anti-NSDHL antibody [EPR14490] (ab190353) 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 - ab190353 observed at 38 kDa. Red - loading control ab7291 observed at 50 kDa.

       ab190353 Anti-NSDHL antibody [EPR14490]  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. ab190353 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSDHL antibody [EPR14490] (ab190353)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSDHL antibody [EPR14490] (ab190353)

      Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of endometrium tissue labeling NSDHL with ab190353 at 1/100 dilution. Secondary antibody HRP Polymer for Rabbit IgG prediluted. Counter stain Hematoxylin. Negative control also shown.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSDHL antibody [EPR14490] (ab190353)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSDHL antibody [EPR14490] (ab190353)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NSDHL with ab190353 at 1/100 dilution. Secondary antibody HRP Polymer for Rabbit IgG prediluted. Counter stain Hematoxylin. Negative control also shown.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-NSDHL antibody [EPR14490] (ab190353)
      Immunocytochemistry/ Immunofluorescence - Anti-NSDHL antibody [EPR14490] (ab190353)

      Immunofluorescence analysis of U87-MG cells, labeling NSDHL (red) with ab190353 at 1/50 dilution. Alexa Fluor®555-conjugated goat anti-rabbit IgG was used as a secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).

    • Western blot - Anti-NSDHL antibody [EPR14490] (ab190353)
      Western blot - Anti-NSDHL antibody [EPR14490] (ab190353)
      All lanes : Anti-NSDHL antibody [EPR14490] (ab190353) at 1/10000 dilution

      Lane 1 : A431 cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : Human fetal brain lysate

      Lysates/proteins at 20 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: 38 kDa why is the actual band size different from the predicted?

    References (1)

    Publishing research using ab190353? Please let us know so that we can cite the reference in this datasheet.

    ab190353 has been referenced in 1 publication.

    • Ricco N  et al. Mevalonate pathway activity as a determinant of radiation sensitivity in head and neck cancer. Mol Oncol 13:1927-1943 (2019). PubMed: 31225926

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