Anti-Ninein antibody - Centrosome Marker (ab4447)

Overview

  • Product name
    Anti-Ninein antibody - Centrosome Marker
    See all Ninein primary antibodies
  • Description
    Rabbit polyclonal to Ninein - Centrosome Marker
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Fusion protein containing amino acids 32-506 of murine Ninein

  • Positive control
    • This antibody gave a positive signal in the following cell lines: HeLa; MCF7.
  • General notes


    The centrosomal protein ninein has been identified as a microtubule minus end capping and centriole anchoring protein, but its true physiological function remains to be determined. At mitosis, ninein has been observed in association with the mitotic spindle as well as at the centrosomes.

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
  • Concentration information loading...
  • Purity
    Protein A purified
  • Primary antibody notes
    The centrosomal protein ninein has been identified as a microtubule minus end capping and centriole anchoring protein, but its true physiological function remains to be determined. At mitosis, ninein has been observed in association with the mitotic spindle as well as at the centrosomes.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab4447 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 Use at an assay dependent concentration. PubMed: 28469279
WB 1/500. Detects a band of approximately 249 kDa (predicted molecular weight: 249 kDa).
ICC/IF 1/500.

Target

Anti-Ninein antibody - Centrosome Marker images

  • IF using ab4447 to detect Ninein in MCF7 cells (human).

    The left hand panels are DAPI stained nuclei. The right hand panels are the same nuclei stained for Ninein.

    ab4447 was used at  1/1000.

    Ninein shows nuclear and centrosome staining, which is expected from the literature.

  • ICC/IF image of ab4447 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4447, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    ab4447 shows the characteristic nuclear and centrosome staining for Ninein.

References for Anti-Ninein antibody - Centrosome Marker (ab4447)

This product has been referenced in:
  • Magescas J  et al. Spindle pole cohesion requires glycosylation-mediated localization of NuMA. Sci Rep 7:1474 (2017). IHC-P ; Mouse . Read more (PubMed: 28469279) »
  • Jao LE  et al. A role for Gle1, a regulator of DEAD-box RNA helicases, at centrosomes and basal bodies. Mol Biol Cell 28:120-127 (2017). Read more (PubMed: 28035044) »

See all 25 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mIMCD-3 kidney epithelial cells)
Specification
mIMCD-3 kidney epithelial cells
Fixative
Methanol
Permeabilization
Yes - NP40
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Mr. Benjamin Tyrrell

Verified customer

Submitted Apr 13 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (ARPE-19 cells)
Specification
ARPE-19 cells
Fixative
Methanol
Permeabilization
Yes - NP40
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Mr. Benjamin Tyrrell

Verified customer

Submitted Apr 13 2010

Thank you for your enquiry. The investigator who provided the images for our datasheet for ab4447 used this protocol: Fix in 3.5% paraformaldehyde (in PBS with final pH6.9). 10 minutes should be sufficient. Permeablize in 0.2% TX100 in KB buffe...

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Thank you for your enquiry. Further to our conversation this morning I have discovered that the immunogen used in this case was a fusion protein containing amino acids 32-506 of murine Ninein. I have retrieved the sequence from the EXPASY database for ...

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Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab4447. Without knowing more details regarding your protocol (dilutions tried? what was was your secondary antibody? what type of detection method are you using?...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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