Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)

Reviews (2)Q&A (3)References (25)

Overview

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

    Fusion protein corresponding to Mouse Ninein/NIN aa 1-550.

  • 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

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)
    Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)Gordon Chan, University of Alberta

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)
    Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)

    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

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

Publishing research using ab4447? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ninein antibody - Centrosome Marker

 Inconclusive
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
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
Read More

Mr. Benjamin Tyrrell

Verified customer

Submitted Apr 13 2010

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ninein antibody - Centrosome Marker

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
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
Read More

Mr. Benjamin Tyrrell

Verified customer

Submitted Apr 13 2010

Question

The customer would like to know the fixation protocol used by the researcher who provided the b&w image on the datasheet.

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Answer

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 buffer(150mM NaCl, 10mM TrispH7.8, 0.1%BSA) for 5 minute and wash in KB buffer (minus TX100) for another 5 minutes before primary antibody incubation, 1/1000. Please do not hesitate to contact us if you have additional questions.

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Question

Mette would like information about the immunogen.

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Answer

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 your convenience: ELTDLCHVLCLEDVGPVLQQTLLQDNLLGRVHFDQFKEALILILSRTLSSEEHFEES DCSPEAQPKYVRGGKRYGRRSLPEFQESGEEIEEVTVLEPLEEEARSSPIPAGDCGE HWKTQRSEEYEAEGQLRFWNPDDLNASHGGSCPPPDWIEEKLQEVCEDLGITRDGHL NRKKLVSICEQYGLQNVDGAMLEEVFLSLDPDGTMSVEDFFYGLFKTGKSLTPSAST PYRQLKRHLSMQSFDESGRRTATSSAMTSTIGFRVFSCLDDGMGQASVERILDTWQE EGIENSQEILKALDFSLDGNINLTELTLALENELLVTKNGIHQAALASFKAEIRHLL ERVDQVVREKEKLRSDLDKAEKLKSLMASEVDDHHAAIERRNEYNLRKLDEEYKERI AALKNELRQEREQMLQQVGKQRVELEQEIQKAKTEENYIRDRLALSLKENNRLETEL LENAEKLAEYESLTQKLQR I hope this information helps, please do not hesitate to contact me should you require further assistance.

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM High background SAMPLE cultured cell lines FIXATION OF SAMPLE tried methanol and 4%pfa BLOCKING CONDITIONS tried BSA HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? tried both methanol and 4%pfa fixation, with high bckgrnd both times

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Answer

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? what was the batch that you received? what type of cells are you using?), I would like to make a few suggestions. Try increasing the blocking incubation period. Also, the concentration of ab4447 may be too high; make sure to titrate the antibody. We do recommend using ab4447 at a concentration of 1:1000. Your secondary antibody may be binding non-specifically; make sure to a secondary control (no primary antibody), and also decrease the concentration of the secondary antibody. Please let me know if you have any additional questions.

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