Anti-Eg5 antibody [EPR23276-52] (ab254298)

Rabbit Recombinant Monoclonal Eg5 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human samples. Cited in 4 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23276-52] (AB254298), expandable thumbnail

Publications

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

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P	IHC-Fr
Human	Tested	Tested	Tested	Tested	Tested	Not recommended
Mouse	Tested	Tested	Tested	Tested	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes -
Species Human	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/600	Notes -
Species Human	Dilution info 1/600	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
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	Dilution info	Notes
Species Rat, Mouse, Human	Dilution info -	Notes -

Associated Products

Select an associated product type

7 products for Alternative Product

Target data

See full target information KIF11

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 Mouse, Human samples. Cited in 4 publications.

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.
Clone number: EPR23276-52
Purification technique: Affinity purification Protein A
Concentration

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:
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12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Eg5 with 254298 at 1/1/4000 (0.17μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in spermatocytes of human testis is observed.
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Eg5 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug with ab254298 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254298 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10 ug.
Lane 2: ab254298 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254298 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
Fresh lysates were used in this IP.
All lanes: Immunoprecipitation - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Predicted band size: 119 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling Eg5 with ab254298 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 2 μg/ml dilution (Green). Confocal image showing cytoplasmic and spindle (arrow) staining in NIH/3T3 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: 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 2 μg/ml dilution.
Immunoprecipitation - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Eg5 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab254298 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254298 at 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 ug.
Lane 2: ab254298 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254298 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Fresh lysates were used in this IP.
All lanes: Immunoprecipitation - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Predicted band size: 119 kDa
Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
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 may correspond to degradation.
Exposure time: 48 seconds.
All lanes: Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298) 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 1/100000 dilution
Predicted band size: 119 kDa
Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labeling Eg5 with ab254298 at 1/600 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A 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.
Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Eg5 with ab254298 at 1/600 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A 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.
Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Eg5 with ab254298 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 2 μg/ml dilution (Green). Confocal image showing cytoplasmic and spindle (arrow) staining in HeLa cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: 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 2 μg/ml dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Eg5 with ab254298 at 1/4000 (0.17μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Scattered cytoplasmic staining in human breast cancer cells is observed (PMID:29181100).
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Eg5 with 254298 at 1/4000 (0.17μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in human tonsil is observed (PMID:25277178).
The section was incubated with ab254298 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID:23857769; 27492783).
Fresh lysates were tested in this WB.
The minor bands beneath the target band in lane 1 and lane 4 may correspond to degradation.
Exposure time: 15 seconds.
All lanes: Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298) at 1/1000 dilution
Lane 1: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg
Lane 2: NCI-H1975 (human adenocarcinoma lung epithelial cell) at 20 µg
Lane 3: MCF7 (human breast adenocarcinoma epithelial cell) at 20 µg
Lane 4: 4T1 (mouse mammary gland carcinoma epithelial cell) 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
Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Mouse brain (PMID: 17974955).
The minor bands beneath the target band may correspond to degradation.
Exposure time: 48 seconds.
All lanes: Western blot - Anti-Eg5 antibody [EPR23276-52] (ab254298) at 1/1000 dilution
Lane 1: Mouse testis tissue lysate at 20 µg
Lane 2: Mouse brain tissue 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