Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

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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 [EPR19993] to Niemann Pick C2
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR19993] to Niemann Pick C2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Niemann Pick C2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P61916

  • Positive control

    • WB: Human fetal kidney and spleen lysates; HEK-293T, HepG2, C6, RAW 264,7, PC-12 and NIH/3T3 whole cell lysates; mouse spleen lysate; rat kidney lysate. IHC-P: Human endometrium and endometrium cancer tissues; mouse testis tissue; rat kidney tissue.

  • 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

Our Abpromise guarantee covers the use of ab207158 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
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

ab93684 can be used for antigen retrieval buffer.
WB 1/1000. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).

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

    see all

  • 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] (ab207158)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    All lanes : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) 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 - ab207158 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab207158 Anti-Niemann Pick C2 antibody [EPR19993] 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. ab207158 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

    Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human endometrium cancer 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) (ab97051) at 1/500 dilution.

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

  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Lanes 1 & 3 : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/1000 dilution
    Lane 2 : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/10000 dilution

    Lane 1 : Human fetal kidney lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 3 : Human spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG 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?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 3 seconds; Lane 3: 30 seconds.

    NPC2 can be glycosylated, the WB pattern is consistent with references PMID:16374838 and PMID:22183894.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

    Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human endometrium 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) (ab97051) at 1/500 dilution.

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

  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    All lanes : Anti-Niemann Pick C2 antibody [EPR19993] (ab207158) at 1/1000 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
    Lane 5 : Mouse spleen lysate
    Lane 6 : Rat kidney 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?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3/5: 3 minutes; Lane 4/6: 10 seconds.

    NPC2 can be glycosylated, the WB pattern is consistent with references PMID:16374838 and PMID:22183894.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse testis 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) (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] (ab207158)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993] (ab207158)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Niemann Pick C2 with ab207158 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.  Cytoplasmic staining on rat kidney 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) (ab97051) at 1/500 dilution.

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

References (0)

Publishing research using ab207158? Please let us know so that we can cite the reference in this datasheet.

ab207158 has not yet been referenced specifically in any publications.

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