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Rabbit Recombinant Monoclonal Eg5 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (AB254299), expandable thumbnail
  • Western blot - Anti-Eg5 antibody [EPR23277-60] (AB254299), expandable thumbnail
  • Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (AB254299), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-60] (AB254299), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-60] (AB254299), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-PFlow Cyt
Human
Tested
Tested
Tested
Tested
Tested
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/30
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/50
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/500
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Rat, Mouse, Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

7 products for Alternative Product

Target data

Function

Motor protein required for establishing a bipolar spindle and thus contributing to chromosome congression during mitosis (PubMed:19001501, PubMed:37728657). Required in non-mitotic cells for transport of secretory proteins from the Golgi complex to the cell surface (PubMed:23857769).

Alternative names

EG5, KNSL1, TRIP5, KIF11, Kinesin-like protein KIF11, Kinesin-like protein 1, Kinesin-like spindle protein HKSP, Kinesin-related motor protein Eg5, Thyroid receptor-interacting protein 5, TR-interacting protein 5, TRIP-5

Recommended products

Rabbit Recombinant Monoclonal Eg5 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR23277-60
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The Eg5 protein also known scientifically as kinesin family member 11 (KIF11) plays an important role in cell mitosis. It functions mechanically as a plus-end directed motor involved in the separation of centrosomes during the early stage of cell division. Eg5 with an approximate mass of 116 kDa is responsible for the proper alignment and segregation of spindle fibers. The protein is expressed in rapidly dividing cells particularly within tissues with high turnover rates such as bone marrow and epithelium.

Biological function summary

The motor function of Eg5 is indispensable for the mitotic spindle organization. It provides the force necessary to push microtubules in opposite directions. Eg5 functions as part of a complex cellular machinery that includes proteins like dynein and other kinesins. By controlling spindle bipolarity it ensures accurate chromosome segregation during cell division preventing errors that could lead to aneuploidy.

Pathways

The action of Eg5 is fundamental in mitotic pathways including the spindle assembly checkpoint pathway. It interacts with proteins such as cyclin-dependent kinases (CDKs) which regulate its activity to ensure proper cell cycle progression. The protein's role is integral in facilitating the dynamics of the metaphase spindle checkpoint guaranteeing cell cycle fidelity.

Associated diseases and disorders

Eg5 is linked to certain types of cancers where there is abnormal cell division. The overexpression or dysregulation of Eg5 can contribute to tumorigenesis. Additionally Eg5 interacts with proteins involved in cancer such as p53 a tumor suppressor protein directly influencing cellular proliferation and survival. Researchers target Eg5 in the development of anticancer therapies to inhibit its activity aiming to obstruct malignant cell proliferation.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Eg5 with ab254299 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID:25277178) is observed.

    The section was incubated with ab254299 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time:

    Lanes 1-2: 15 seconds;
    Lane 3: 5.5 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID:23857769; 27492783).

    Fresh lysates were tested in this WB.

    All lanes: Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299) at 1/1000 dilution

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 3: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 119 kDa

    Observed band size: 130 kDa

  • Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Eg5 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate with ab254299 at 1/30 dilution. Western blot was performed on the immunoprecipitate using 254299 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg

    Lane 2: ab254299 IP in HeLa whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254299 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds

    The minor bands beneath the target band are probably caused by degradation.

    All lanes: Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Predicted band size: 119 kDa

    Observed band size: 130 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell line) cells labelling Eg5 with ab254299 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Eg5 with ab254299 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Eg5 with ab254299 at 1/500 dilution (1.18 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID:25277178) is observed.

    The section was incubated with ab254299 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 15 seconds

    The expression profile observed is consistent with what has been described in the literature (PMID:23857769; 27492783).

    Negative control: Human lung (PMID: 27279560).

    The minor bands beneath the target band are probably caused by degradation.

    All lanes: Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299) at 1/1000 dilution

    Lane 1: Human testis tissue lysate at 20 µg

    Lane 2: Human tonsil tissue lysate at 20 µg

    Lane 3: Human lung tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

    Predicted band size: 119 kDa

    Observed band size: 130 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)

    Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Eg5 with ab254299 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Cytoplasmic staining in human tonsil (PMID:25277178) is observed. The section was incubated with ab254299 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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