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

Reviews (2)Q&A (3)References (30)
Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ninein/NIN antibody - Centrosome Marker (ab4447)

Key features and details

  • Rabbit polyclonal to Ninein/NIN - Centrosome Marker
  • Suitable for: ICC/IF
  • Reacts with: Human
  • Isotype: IgG

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 & Species

    Application Species
    ICC/IF
    Human
    See all applications and species data

  • Immunogen

    Fusion protein corresponding to Human Ninein/NIN. The region used shows good homology to Mouse Ninein/NIN aa 1-550.

  • 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.

    This product was previously labelled as Ninein.

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

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
All applications
Mouse
Application Abreviews Notes
ICC/IF (2)
1/500.
Notes
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/NIN in MCF7 cells (human).

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

    ab4447 was used at  1/1000.

    Ninein/NIN 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/NIN.

     

References (30)

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

ab4447 has been referenced in 30 publications.

  • Aydin ÖZ  et al. Acute inhibition of centriolar satellite function and positioning reveals their functions at the primary cilium. PLoS Biol 18:e3000679 (2020). PubMed: 32555591
  • Ogungbenro YA  et al. Centrobin controls primary ciliogenesis in vertebrates. J Cell Biol 217:1205-1215 (2018). PubMed: 29440264
  • Shi CH  et al. Disrupted structure and aberrant function of CHIP mediates the loss of motor and cognitive function in preclinical models of SCAR16. PLoS Genet 14:e1007664 (2018). PubMed: 30222779
  • Rowlison T  et al. Deciphering the mechanisms involving cenexin, ninein and centriolin in sperm maturation during epididymal transit in the domestic cat. Reprod Domest Anim 52 Suppl 2:193-196 (2017). PubMed: 27807895
  • 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). PubMed: 28035044
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

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

 Inconclusive
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mIMCD-3 kidney epithelial cells)
Permeabilization
Yes - NP40
Specification
mIMCD-3 kidney epithelial cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Methanol
Read More

Mr. Benjamin Tyrrell

Verified customer

Submitted Apr 13 2010

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

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (ARPE-19 cells)
Permeabilization
Yes - NP40
Specification
ARPE-19 cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Methanol
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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