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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

Reviews (1) Submit a question References (1)
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
  • Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19993-145-1] to Niemann Pick C2
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Niemann Pick C2 antibody [EPR19993-145-1]
    See all Niemann Pick C2 primary antibodies

  • Description

    Rabbit monoclonal [EPR19993-145-1] to Niemann Pick C2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal kidney and spleen lysates; Mouse kidney lysate; Rat kidney and spleen lysates; HEK-293T, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human epididymis and testis tissues; Mouse and rat kidney tissues. ICC/IF: HepG2 and NIH/3T3 cells. Flow Cyt (intra): HepG2 and NIH/3T3 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab218192 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
Flow Cyt (Intra)
1/70.
WB (1)
1/2000. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
Notes
Flow Cyt (Intra)
1/70.
WB
1/2000. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.

Target

  • Function

    May be involved in the regulation of the lipid composition of sperm membranes during the maturation in the epididymis.

  • Tissue specificity

    Epididymis.

  • Involvement in disease

    Defects in NPC2 are the cause of Niemann-Pick disease type C2 (NPDC2) [MIM:607625]. A lysosomal storage disorder that affects the viscera and the central nervous system. It is due to defective intracellular processing and transport of low-density lipoprotein derived cholesterol. It causes accumulation of cholesterol in lysosomes, with delayed induction of cholesterol homeostatic reactions. Niemann-Pick disease type C2 has a highly variable clinical phenotype. Clinical features include variable hepatosplenomegaly and severe progressive neurological dysfunction such as ataxia, dystonia and dementia. The age of onset can vary from infancy to late adulthood.

  • Sequence similarities

    Belongs to the NPC2 family.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    • Alternative names

      • EDDM1 antibody
      • Epididymal protein 1 antibody
      • Epididymal secretory protein antibody
      • Epididymal secretory protein E1 antibody
      • hE1 antibody
      • Human epididymis-specific protein 1 antibody
      • Niemann-Pick disease type C2 antibody
      • Niemann-Pick disease type C2 protein antibody
      • NPC intracellular cholesterol transporter 2 antibody
      • NPC2 antibody
      • NPC2_HUMAN antibody
      • Tissue specific secretory protein antibody
      see all

    Images

    • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 2 : NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole 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: 17 kDa
      Observed band size: 16-18 kDa why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab218192 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab218192 Anti-Niemann Pick C2 antibody [EPR19993-145-1] was shown to specifically react with Niemann Pick C2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266749 (knockout cell lysate ab258079) was used. Wild-type and Niemann Pick C2 knockout samples were subjected to SDS-PAGE. ab218192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Niemann Pick C2 with ab218192 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunohistochemical analysis of paraffin-embedded human epididymis tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human epididymis [PMID: 8924505]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

    • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Niemann Pick C2 with ab218192 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) at 1/5000 dilution

      Lane 1 : Human fetal kidney lysate
      Lane 2 : Mouse kidney lysate
      Lane 3 : Rat kidney lysate
      Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 5 : Rat spleen lysate
      Lane 6 : Human spleen lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 17 kDa
      Observed band size: 16-18 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lane 1-4: 4 seconds; Lane 5/6: 15 seconds.

      Niemann Pick C2 can be glycosylated, the WB pattern is consistent with references PMID: 16374838; PMID: 22183894. 

    • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) at 1/2000 dilution

      Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
      Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 17 kDa
      Observed band size: 16-18 kDa why is the actual band size different from the predicted?


      Exposure time: 10 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Niemann Pick C2 can be glycosylated, the WB pattern is consistent with references PMID:16374838; PMID: 22183894.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on human testis [PMID: 8924505]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      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-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on mouse kidney tubules [PMID: 24147030]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      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-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on rat kidney tubules [PMID: 24147030]. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

    • Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Niemann Pick C2 with ab218192 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Niemann Pick C2 with ab218192 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)
      Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192)

    References (1)

    Publishing research using ab218192? Please let us know so that we can cite the reference in this datasheet.

    ab218192 has been referenced in 1 publication.

    • Heybrock S  et al. Lysosomal integral membrane protein-2 (LIMP-2/SCARB2) is involved in lysosomal cholesterol export. Nat Commun 10:3521 (2019). PubMed: 31387993

    Customer reviews and Q&As

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    Western blot abreview for Anti-Niemann Pick C2 antibody [EPR19993-145-1]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Fibroblast)
    Gel Running Conditions
    Reduced Denaturing (10% Bis Tris - Mes buffer Migration)
    Loading amount
    20 µg
    Treatment
    MG132 1uM 24h
    Specification
    Fibroblast
    Blocking step
    Specific blocking buffer for fluorescence revelation as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: RT°C
    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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    Verified customer

    Submitted Aug 22 2022

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