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

Reviews (2)Q&A (3)References (31)
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

概述

  • 产品名称

    Anti-Ninein/NIN抗体- Centrosome Marker
    参阅全部 Ninein/NIN 一抗

  • 描述

    兔多克隆抗体to Ninein/NIN - Centrosome Marker

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IFmore details

  • 种属反应性

    与反应: Human
    预测可用于: Mouse

  • 免疫原

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

  • 阳性对照

    • ICC/IF: HeLa cells

  • 常规说明

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid

  • 存放说明

    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.

  • 存储溶液

    pH: 7.4
    Preservative: 0.02% Sodium azide
    Constituents: 98.98% PBS, 1% BSA
  • Concentration information loading...

  • 纯度

    Protein A purified

  • Primary antibody说明

    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.

  • 克隆

    多克隆

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4447于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF (2)
1/500.
说明
ICC/IF
1/500.

靶标

图片

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

    ab4447 staining Ninein in HeLa cells. The cells were fixed with 4% PFA (10 mins), 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 ab4447 (shown in green) at 0.1µg/ml and ab7291 (shown in red), Mouse monoclonal to alpha Tubulin, at 1µg/ml. The secondary antibodies used were ab150081, Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L), and ab150120, Alexa Fluor® 594 Goat Anti-Mouse IgG (H&L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • 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.

文献 (31)

发表研究结果有使用 ab4447？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab4447 被引用在 31 文献中.

  • 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
  • Tona Y & Wu DK Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2. Elife 9:N/A (2020). PubMed: 32965215
  • 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
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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
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.

Mr. Benjamin Tyrrell

Verified customer

提交于 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
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.

Mr. Benjamin Tyrrell

Verified customer

提交于 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

Read More
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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