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    niemann-pick-c1-antibody-epr5209-ab134113.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

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Reviews (1)Q&A (1)References (30)

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Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Flow Cytometry (Intracellular) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • OI-RD Scanning - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
  • Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5209] to Niemann Pick C1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free

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Recombinant Human Niemann Pick C1 protein (ab114306)
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Overview

  • Product name

    Anti-Niemann Pick C1 antibody [EPR5209]
    See all Niemann Pick C1 primary antibodies
  • Description

    Rabbit monoclonal [EPR5209] to Niemann Pick C1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Niemann Pick C1 aa 1250 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: O15118

  • Positive control

    • WB: Wild-type HAP1 whole cell lysate; HEK-293, HepG2, THP-1, PC-3, 3T3L1, L6 and HEK-293T cell lysates; Rat liver and brain lysates. IHC-P: Human kidney and liver tissues; Mouse liver tissue; Rat cerebellum tissue. ICC/IF: Neuro-2a cells; SH-SY5Y control and PADK treated cells. Flow Cyt (intra): Neuro-2a 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 4.90 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5209
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Transporters
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Coat Proteins
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Cardiovascular
    • Atherosclerosis
    • Lipoprotein metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipoprotein metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Fatty acids
    • Metabolism
    • Types of disease
    • Heart disease
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation

Associated products

  • Alternative Versions

    • APC Anti-Niemann Pick C1 antibody [EPR5209] (ab223985)
    • Anti-Niemann Pick C1 antibody [EPR5209] - BSA and Azide free (ab224268)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Rat brain normal tissue lysate - membrane extract (ab29473)
  • Recombinant Protein

    • Recombinant Human Niemann Pick C1 protein (ab114306)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab134113 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)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/2000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 142 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/70.
Notes
Flow Cyt (Intra)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/2000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 142 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/70.

Target

  • Function

    Involved in the intracellular trafficking of cholesterol. May play a role in vesicular trafficking in glia, a process that may be crucial for maintaining the structural and functional integrity of nerve terminals.
  • Involvement in disease

    Defects in NPC1 are the cause of Niemann-Pick disease type C1 (NPDC1) [MIM:257220]. 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 C1 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. An allelic variant of Niemann-Pick disease type C1 is found in people with Nova Scotia ancestry. Patients with the Nova Scotian clinical variant are less severely affected.
  • Sequence similarities

    Belongs to the patched family.
    Contains 1 SSD (sterol-sensing) domain.
  • Domain

    A cysteine-rich N-terminal domain and a C-terminal domain containing a di-leucine motif necessary for lysosomal targeting are critical for mobilization of cholesterol from lysosomes.
  • Post-translational
    modifications

    Glycosylated.
  • Cellular localization

    Late endosome membrane. Lysosome membrane.
  • Target information above from: UniProt accession O15118 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4864 Human
    • Entrez Gene: 18145 Mouse
    • Entrez Gene: 266732 Rat
    • Omim: 607623 Human
    • SwissProt: O15118 Human
    • SwissProt: O35604 Mouse
    • Unigene: 464779 Human
    • Unigene: 715623 Human
    • Unigene: 3484 Mouse
    see all
  • Alternative names

    • Niemann Pick C1 protein precursor antibody
    • Niemann Pick disease, type C1 antibody
    • Niemann-Pick C1 protein antibody
    • NPC antibody
    • NPC1 antibody
    • NPC1_HUMAN antibody
    see all

Images

  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : NPC1 (Niemann Pick C1) knockout HAP1 whole cell lysate
    Lane 3 : HEK293 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 142 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab134113 observed at 180 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab134113 was shown to specifically react with Niemann Pick C1 in wild-type HAP1 cells as signal was lost in NPC1 (Niemann Pick C1) knockout cells. Wild-type and NPC1 (Niemann Pick C1) knockout samples were subjected to SDS-PAGE. Ab134113 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Niemann Pick C1 with ab134113 at 1/70 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500(green). Confocal image showing cytoplasmic staining on Neuro-2a cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120(AlexaFluor®594 Goat anti-Mouse secondary) at 1/500  (red).

    The negative controls are as follows:
    -ve control 1 – ab134113 at 1/70 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    Immunohistochemical staining of paraffin embedded human liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Flow Cytometry (Intracellular) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Flow Cytometry (Intracellular) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    ab134113 staining Niemann Pick C1 in Neuro-2a (mouse neuroblastoma cell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody. 

    Isoytype control: Rabbit monoclonal IgG (Black) 

    Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue) 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)Schultz et al Nat Commun. 2018; 9: 3671. Published online 2018 Sep 10. doi: 10.1038/s41467-018-06115-2

    I1061T NPC1 traffics to autophagosomes 

    I1061T NPC1 fibroblasts were treated with vehicle or 100 nM bafilomycin A1 (Baf) for 24 h. Fixed cells were stained for LC3 (red), NPC1 (green), and DAPI (blue) then imaged by confocal microscopy. Co-localization is indicated by yellow color in the merged image. Scale bar = 25 μm.

    Niemann Pick C1 (NPC1) was detected using ab134113 at 1/200 dilution.

    From Figure 4b of Schultz et al.

    Shultz et al Nat Commun. 2018; 9: 3671.Published online 2018 Sep 10. doi: 10.1038/s41467-018-06115-2

    Reproduced under the Creative Commons Licence http://creativecommons.org/licenses/by/4.0/.

  • Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)Image from Cermak S et al., PLoS One. 2016;11(11):e0167428. Fig 3.; doi: 10.1371/journal.pone.0167428. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Cathepsin B/L inhibition causes NPC disease-like cholesterol accumulation in SH-SY5Y cells.

    Confocal microscopy of SH-SY5Y control and PADK treated cells. Cholesterol (filipin staining, white) and NPC1 (ab134113; green).

  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)

    Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
    Lane 2 : THP-1 (human monocytic leukemia cell line) cell lysate
    Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
    Lane 4 : PC-3 (human prostate adenocarcinoma cell line) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 142 kDa
    Additional bands at: 180 kDa (possible glycosylated form)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)

    Lane 1 : 3T3-L1 cell lysate
    Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
    Lane 3 : Rat tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 142 kDa
    Additional bands at: 180 kDa (possible glycosylated form)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    Immunohistochemical staining of paraffin embedded mouse liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    Immunohistochemical staining of paraffin embedded rat cerebellum with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Western blot - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution (unpurified)

    Lane 1 : 3T3 L1 cell lysate
    Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
    Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
    Lane 4 : THP-1 (human monocytic leukemia cell line) cell lysate
    Lane 5 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
    Lane 6 : PC-3 (human prostate adenocarcinoma cell line) cell lysate
    Lane 7 : Rat liver lysate
    Lane 8 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 142 kDa
    Observed band size: 180 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

    Immunohistochemical analysis of paraffin embedded human kidney tissue labelling Niemann Pick C1 with unpurified ab134113 at 1/50.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    OI-RD Scanning - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
    Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (30)

Publishing research using ab134113? Please let us know so that we can cite the reference in this datasheet.

ab134113 has been referenced in 30 publications.

  • Stewart CM  et al. Ebola virus triggers receptor tyrosine kinase-dependent signaling to promote the delivery of viral particles to entry-conducive intracellular compartments. PLoS Pathog 17:e1009275 (2021). PubMed: 33513206
  • Sharma C  et al. Antioxidant functions of DHHC3 suppress anti-cancer drug activities. Cell Mol Life Sci 78:2341-2353 (2021). PubMed: 32986127
  • Sudhakar JN  et al. Lumenal Galectin-9-Lamp2 interaction regulates lysosome and autophagy to prevent pathogenesis in the intestine and pancreas. Nat Commun 11:4286 (2020). PubMed: 32855403
  • Haldar S  et al. Precise Triggering and Chemical Control of Single-Virus Fusion within Endosomes. J Virol 95:N/A (2020). PubMed: 33115879
  • Takadate Y  et al. Niemann-Pick C1 Heterogeneity of Bat Cells Controls Filovirus Tropism. Cell Rep 30:308-319.e5 (2020). PubMed: 31940478
View all Publications for this product

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Niemann Pick C1 antibody [EPR5209]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (RPE1 cell line)
Specification
RPE1 cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Formaldehyde
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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Submitted Mar 10 2021

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