Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)

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 - BSA and Azide free
  • Suitable for: Flow Cyt, ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free
    See all Niemann Pick C2 primary antibodies

  • Description

    Rabbit monoclonal [EPR19993-145-1] to Niemann Pick C2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Rat
    WB
    Human
    See all applications and species data

  • 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. Flow Cyt: HepG2 and NIH/3T3 cells.

  • General notes

    Ab251560 is the carrier-free version of ab218192. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab251560 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    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?



    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

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

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

  • Immunocytochemistry - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Immunocytochemistry - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)

    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab251560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)

    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    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 - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Immunocytochemistry - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)

    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab251560)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    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


    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

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

  • Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    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?



    This data was developed using ab218192, the same antibody clone in a different buffer formulation.

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    This data was developed using ab218192, the same antibody clone in a different buffer formulation.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] - BSA and Azide free (ab251560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    This data was developed using ab218192, the same antibody clone in a different buffer formulation.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] - BSA and Azide free (ab251560)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    This data was developed using ab218192, the same antibody clone in a different buffer formulation.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 - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Flow Cytometry - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    This data was developed using ab218192, the same antibody clone in a different buffer formulation.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 - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    Flow Cytometry - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)
    This data was developed using ab218192, the same antibody clone in a different buffer formulation.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] - BSA and Azide free (ab251560)
    Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (ab251560)

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