Recombinant Anti-Eg5 antibody [EPR23277-7] (ab272220)

Product name

Anti-Eg5 antibody [EPR23277-7]
See all Eg5 primary antibodies
Description

Rabbit monoclonal [EPR23277-7] to Eg5
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
Unsuitable for: IHC-Fr or IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human testis, human tonsil, human lung, Jurkat, NCI-H1975, MCF7, 4T1, HeLa, HEK-293, AR42J, mouse testis, rat testis and PC-12, NIH/3T3 lysates. IHC-P: Human tonsil, human breast cancer and mouse testis, Rat testis tissues. ICC/IF: and HeLa, NIH/3T3 cells. Flow Cyt (intra): HeLa cell.
General notes
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 production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23277-7
Isotype

IgG
Research areas

Alternative Versions
- Anti-Eg5 antibody [EPR23277-7] - BSA and Azide free (ab272226)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab272220 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
Flow Cyt (Intra)		1/50.
ICC/IF		1/50.
WB		1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 119 kDa).
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
Flow Cyt (Intra) 1/50.
ICC/IF 1/50.
WB 1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 119 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Application notes

Is unsuitable for IHC-Fr or IP.

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.
Target information above from: UniProt accession P52732 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3832 Human
- Entrez Gene: 16551 Mouse
- Entrez Gene: 171304 Rat
- Omim: 148760 Human
- SwissProt: P52732 Human
- SwissProt: Q6P9P6 Mouse
- Unigene: 8878 Human
- Unigene: 42203 Mouse
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

Western blot - Anti-Eg5 antibody [EPR23277-7] (ab272220)

All lanes : Anti-Eg5 antibody [EPR23277-7] (ab272220) at 1/1000 dilution

Lane 1 : Human testis tissue lysate
Lane 2 : Human tonsil tissue lysate
Lane 3 : Human lung tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 119 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?

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: 70 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Eg5 with ab272220 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in germinal center of human tonsil is observed (PMID:25277178). The section was incubated with ab272220 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.
Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Eg5 with ab272220 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing spindle fibre and cytoplasmic staining in HeLa cell line. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Intracellular flow cytometric analysis of 80% methanol fixed 0.1% Tween-20 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Eg5 with ab272220 at 1/50 dilution (0.1µg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were pre-treated with 20µg/ml RNaseA for 30min to minimize the binding between Propidium Iodide (PI) and RNA. Then intracellularly stained with ab272220 and PI. Propidium Iodide Flow Cytometry Kit (ab139418) was used for RNaseA treatment and PI staining.
Western blot - Anti-Eg5 antibody [EPR23277-7] (ab272220)

All lanes : Anti-Eg5 antibody [EPR23277-7] (ab272220) at 1/1000 dilution

Lane 1 : Mouse testis tissue lysate
Lane 2 : Rat testis tissue lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 119 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?

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

The minor bands beneath the target band may correspond to degradation.

Exposure time: Lanes 1-2: 10 seconds; Lane 3: 3 seconds;Lane 4: 26 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Eg5 with ab272220 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in spermatocytes and spermatogonnia of mouse testis is observed. The section was incubated with ab272220 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.
Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling Eg5 with ab272220 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing spindle fibre and cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor^® 594) was used to counterstain tubulin at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Western blot - Anti-Eg5 antibody [EPR23277-7] (ab272220)

All lanes : Anti-Eg5 antibody [EPR23277-7] (ab272220) at 1/1000 dilution

Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : NCI-H1975 (human adenocarcinoma lung epithelial cell), whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate
Lane 5 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 6 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 7 : AR42J (rat pancreatic tumor epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 119 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?

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

Fresh lysates were used in this WB.

Exposure times: Lane 1: 8 seconds; Lanes 2-7: 48 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-7] (ab272220)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Eg5 with ab272220 at 1/500 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in spermatocytes and spermatogonnia of rat testis is observed. The section was incubated with ab272220 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 [EPR23277-7] (ab272220)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Eg5 with ab272220 at 1/500 dilution 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 ab272220 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.
Anti-Eg5 antibody [EPR23277-7] (ab272220)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab272220? Please let us know so that we can cite the reference in this datasheet.

ab272220 has been referenced in 1 publication.

Li Z et al. KIF11 Serves as an Independent Prognostic Factor and Therapeutic Target for Patients With Lung Adenocarcinoma. Front Oncol 11:670218 (2021). PubMed: 33968780

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Post-translational
modifications