Recombinant Anti-Eg5 antibody [EPR23277-60] (ab254299)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381263-anti-eg5-antibody-epr23277-60-immunohistochemistry-human-tonsil.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23277-60] to Eg5
- Suitable for: Flow Cyt (Intra), IHC-P, IP, ICC/IF, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Eg5 antibody [EPR23277-60]
See all Eg5 primary antibodies -
Description
Rabbit monoclonal [EPR23277-60] to Eg5 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, IP, ICC/IF, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, MCF7 and Jurkat whole cell lysates; Human testis and tonsil tissue lysates. IHC-P: Human tonsil, testis and breast tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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
EPR23277-60 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab254299 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) |
1/500.
|
|
| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| IP |
1/30.
|
|
| ICC/IF |
1/50.
|
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| WB |
1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 119 kDa).
|
| Notes |
|---|
|
Flow Cyt (Intra)
1/500. |
|
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
IP
1/30. |
|
ICC/IF
1/50. |
|
WB
1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 119 kDa). |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 3832 Human
- Omim: 148760 Human
- SwissProt: P52732 Human
- Unigene: 8878 Human
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Alternative names
- EG5 antibody
- HKSP antibody
- KIF11 antibody
see all
Images
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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).
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![Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381265-anti-eg5-antibody-epr23277-60-westernblot-hela-mcf7-jurkat.jpg)
Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299)All lanes : Anti-Eg5 antibody [EPR23277-60] (ab254299) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (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 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381277-anti-eg5-antibody-epr23277-60-immunocytochemistry-hela.jpg)
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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells. 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381260-anti-eg5-antibody-epr23277-60-flow-cytometry-hela.jpg)
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 (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381259-anti-eg5-antibody-epr23277-60-immunoprecipitation-hela.jpg)
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)(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 (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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381262-anti-eg5-antibody-epr23277-60-immunohistochemistry-human-testis.jpg)
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)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381264-anti-eg5-antibody-epr23277-60-westernblot-human-testies-tonsil-lung.jpg)
Western blot - Anti-Eg5 antibody [EPR23277-60] (ab254299)All lanes : Anti-Eg5 antibody [EPR23277-60] (ab254299) 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 Detection Reagent (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 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.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-381261-anti-eg5-antibody-epr23277-60-immunohistochemistry-human-breast.jpg)
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).
-
![Anti-Eg5 antibody [EPR23277-60] (ab254299)](https://www.abcam.com/ps/products/254/ab254299/Images/ab254299-9-benefits-of-recombinant-antibodies.png)
Anti-Eg5 antibody [EPR23277-60] (ab254299)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab254299 has not yet been referenced specifically in any publications.