Anti-Eg5抗体[mAbcam 51976] (ab51976)

概述

性能

应用

Our Abpromise guarantee covers the use of ab51976 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Motor protein required for establishing a bipolar spindle. Blocking of KIF11 prevents centrosome migration and arrest cells in mitosis with monoastral microtubule arrays.
  • 疾病相关Defects in KIF11 are the cause of microcephaly with or without chorioretinopathy, lymphedema, or mental retardation (MCLMR) [MIM:152950]. An autosomal dominant disorder that involves an overlapping but variable spectrum of central nervous system and ocular developmental anomalies. Microcephaly ranges from mild to severe and is often associated with mild to moderate developmental delay and a characteristic facial phenotype with upslanting palpebral fissures, broad nose with rounded tip, long philtrum with thin upper lip, prominent chin, and prominent ears. Chorioretinopathy is the most common eye abnormality, but retinal folds, microphthalmia, and myopic and hypermetropic astigmatism have also been reported, and some individuals have no overt ocular phenotype. Congenital lymphedema, when present, is typically confined to the dorsa of the feet, and lymphoscintigraphy reveals the absence of radioactive isotope uptake from the webspaces between the toes.
  • 序列相似性Belongs to the kinesin-like protein family. BimC subfamily.
    Contains 1 kinesin-motor domain.
  • 翻译后修饰Phosphorylated exclusively on serine during S phase, but on both serine and Thr-926 during mitosis, so controlling the association of KIF11 with the spindle apparatus (probably during early prophase). Phosphorylated upon DNA damage, probably by ATM or ATR.
    A subset of this protein primarily localized at the spindle pole is phosphorylated by NEK6 during mitosis; phosphorylation is required for mitotic function.
  • 细胞定位Cytoplasm. Cytoplasm > cytoskeleton > spindle pole.
  • Information by UniProt
  • 数据库链接
  • 别名
    • EG5 antibody
    • HKSP antibody
    • KIF11 antibody
    • KIF11_HUMAN antibody
    • Kinesin family member 11 antibody
    • Kinesin like protein 1 antibody
    • Kinesin-like protein 1 antibody
    • Kinesin-like protein KIF11 antibody
    • Kinesin-like spindle protein HKSP antibody
    • Kinesin-related motor protein Eg5 antibody
    • KNSL1 antibody
    • MCLMR antibody
    • Thyroid receptor-interacting protein 5 antibody
    • TR-interacting protein 5 antibody
    • TRIP-5 antibody
    • TRIP5 antibody
    see all

Anti-Eg5 antibody [mAbcam 51976] 图像

  • All lanes : Anti-Eg5 antibody [mAbcam 51976] (ab51976) at 5 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Rabbit polyclonal to Mouse IgG - H&L (HRP) at 1/2000 dilution

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 120 kDa
  • ICC/IF image of ab51976 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab51976, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Eg5 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Mouse monoclonal to Eg5 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab51976.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 120kDa: Eg5

Anti-Eg5 antibody [mAbcam 51976] (ab51976)参考文献

This product has been referenced in:
  • Drosopoulos K  et al. APC/C is an essential regulator of centrosome clustering. Nat Commun 5:3686 (2014). Read more (PubMed: 24751481) »
  • Dai J  et al. Studies of haspin-depleted cells reveal that spindle-pole integrity in mitosis requires chromosome cohesion. J Cell Sci 122:4168-76 (2009). ICC/IF ; Human . Read more (PubMed: 19910498) »

See all 2 Publications for this product

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (Spindles assembled in Xenopus laevis cytoplasmic e)
Specification Spindles assembled in Xenopus laevis cytoplasmic e
Fixative Formaldehyde and Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Abcam user community

Verified customer

提交于 Nov 17 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - other (Xenopus laevis and tropicalis cytoplasmic egg extr)
Loading amount 40 µg
Specification Xenopus laevis and tropicalis cytoplasmic egg extr
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 17 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Humman Mammary Epithelial Cells)
Specification Humman Mammary Epithelial Cells
Fixative Methanol
Permeabilization Yes - PBS1x-1%TritonX
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
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Ms. Judit Pampalona-Sala

Verified customer

提交于 Aug 13 2008

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