Overview

  • Product name

  • Description

    Rabbit polyclonal to NDP52
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Chimpanzee, Rhesus monkey, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human NDP52 aa 350 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab68587)

  • Positive control

    • This antibody gave a positive signal in the following Human Lysates: TE 671 Whole Cell, HeLa Whole Cell - Hydroxyurea Treated (48hr, 2µM), Ramos Whole Cell, Placenta Tissue ICC-IF: Hela cells untreated, Hela cells chloroquine treated.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab68588 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
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa).
IP Use at an assay dependent concentration. PubMed: 23209807

Target

  • Function

    May play a role in ruffle formation and actin cytoskeleton organization. Seems to negatively regulate constitutive secretion.
  • Tissue specificity

    Expressed in all tissues tested with highest expression in skeletal muscle and lowest in brain.
  • Cellular localization

    Cytoplasm > perinuclear region. Golgi apparatus. Cytoplasm > cytoskeleton. According to PubMed:7540613, localizes to nuclear dots. According to PubMed:9230084 and PubMed:12869526, it is not a nuclear dot-associated protein but localizes predominantly in the cytoplasm with a coarse-grained distribution preferentially close to the nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen nuclear dot 52 kDa protein antibody
    • CACO2_HUMAN antibody
    • Calcium binding and coiled coil domain 2 antibody
    • Calcium binding and coiled coil domain containing protein 2 antibody
    • Calcium-binding and coiled-coil domain-containing protein 2 antibody
    • CALCOCO 2 antibody
    • CALCOCO2 antibody
    • MGC17318 antibody
    • NDP52 antibody
    • Nuclear domain 10 protein 52 antibody
    • Nuclear domain 10 protein antibody
    • Nuclear domain 10 protein NDP52 antibody
    • Nuclear domain protein 52 antibody
    • Nuclear dot protein 52 antibody
    • OTTHUMP00000218344 antibody
    • OTTHUMP00000218345 antibody
    • OTTHUMP00000218346 antibody
    • OTTHUMP00000218347 antibody
    • OTTHUMP00000218349 antibody
    • OTTHUMP00000218351 antibody
    see all

Images

  • ab68588 staining NDP52 in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 4% paraformaldheyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68588 at 5μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control, at 1/1000 dilution. The secondary antibodies were  ab150081 (colored green) and ab150120 (pseudo-colored red) used at 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.

  • All lanes : Anti-NDP52 antibody (ab68588) at 1 µg/ml

    Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
    Lane 2 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2µM)
    Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 4 : Human placenta tissue lysate - total protein (ab29745)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes
  • IHC image of NDP52 staining in Human Cerebral Cortex FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab68588, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX

References

This product has been referenced in:

  • Ravenhill BJ  et al. The Cargo Receptor NDP52 Initiates Selective Autophagy by Recruiting the ULK Complex to Cytosol-Invading Bacteria. Mol Cell 74:320-329.e6 (2019). Read more (PubMed: 30853402) »
  • Baillet N  et al. Autophagy Promotes Infectious Particle Production of Mopeia and Lassa Viruses. Viruses 11:N/A (2019). Read more (PubMed: 30909570) »
See all 28 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (smooth muscle cells)
Permeabilization
Yes - Triton X-100 0,1%
Specification
smooth muscle cells
Blocking step
Gelatin from cold water fish skin as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde

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Verified customer

Submitted Apr 08 2013

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