Anti-Eg5 (phospho T927) antibody (ab61104)

Overview

  • Product name
    Anti-Eg5 (phospho T927) antibody
    See all Eg5 primary antibodies
  • Description
    Rabbit polyclonal to Eg5 (phospho T927)
  • Specificity
    ab61104 detects endogenous levels of Eg5 only when phosphorylated at threonine 927.
  • Tested applications
    Suitable for: WB, ICC/IF, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic phosphopeptide derived from human Eg5 around the phosphorylation site of threonine 927 (G-T-TP-P-Q).

  • Positive control
    • Human lung carcinoma tissue, extracts from COLO205 cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab61104 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
WB 1/500 - 1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).
ICC/IF 1/500 - 1/1000.
ELISA 1/5000.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Motor protein required for establishing a bipolar spindle. Blocking of KIF11 prevents centrosome migration and arrest cells in mitosis with monoastral microtubule arrays.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the kinesin-like protein family. BimC subfamily.
    Contains 1 kinesin-motor domain.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Cytoplasm. Cytoplasm > cytoskeleton > spindle pole.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • All lanes : Anti-Eg5 (phospho T927) antibody (ab61104) at 1/500 dilution

    Lane 1 : Extracts from COLO205 cells.
    Lane 2 : Extracts from COLO205 cells with the immunising peptide


    Predicted band size : 120 kDa
    Observed band size : 120 kDa
  • ICC/IF image of ab61104 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab61104, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab61104 staining in human bone marrow chronic lymphocytic leukemia formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab61104, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

ab61104 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblast Cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
25 µg
Treatment
150 nM nocodazole for 5.5 hrs
Specification
Mouse Embryonic Fibroblast Cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted May 12 2016

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