Recombinant Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)
![Western blot - Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)](https://www.abcam.com/ps/products/251/ab251203/Images/ab251203-395232-anti-bub1-antibody-epr18947-western-blot-hela-lysates.jpg "Western blot - Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18947] to Bub1 - BSA and Azide free
- Suitable for: ICC, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Bub1 antibody [EPR18947] - BSA and Azide free
See all Bub1 primary antibodies -
Description
Rabbit monoclonal [EPR18947] to Bub1 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody shows stronger signal in mouse and rat testis tissues but weaker in the human testis.
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Tested applications
Suitable for: ICC, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human Bub1 aa 1-200. The exact sequence is proprietary.
Database link: O43683 -
General notes
ab251203 is the carrier-free version of ab195268 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
Ab251203 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
Our in house IHC testing showed positive staining in testis tissue ONLY. Other tissues were negative.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Clonality
Monoclonal -
Clone number
EPR18947 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab251203 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 |
|---|---|---|
| ICC | Use at an assay dependent concentration. | |
| WB | Use at an assay dependent concentration. Detects a band of approximately 122 kDa (predicted molecular weight: 122 kDa). | |
| IHC-P | Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Our in house IHC testing showed positive staining in testis tissue ONLY. Other tissues were negative.
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Target
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Function
Serine/threonine-protein kinase that performs 2 crucial functions during mitosis: it is essential for spindle-assembly checkpoint signaling and for correct chromosome alignment. Has a key role in the assembly of checkpoint proteins at the kinetochore, being required for the subsequent localization of CENPF, BUB1B, CENPE and MAD2L1. Required for the kinetochore localization of PLK1. Plays an important role in defining SGOL1 localization and thereby affects sister chromatid cohesion. Acts as a substrate for anaphase-promoting complex or cyclosome (APC/C) in complex with its activator CDH1 (APC/C-Cdh1). Necessary for ensuring proper chromosome segregation and binding to BUB3 is essential for this function. Can regulate chromosome segregation in a kinetochore-independent manner. Can phosphorylate BUB3. The BUB1-BUB3 complex plays a role in the inhibition of APC/C when spindle-assembly checkpoint is activated and inhibits the ubiquitin ligase activity of APC/C by phosphorylating its activator CDC20. This complex can also phosphorylate MAD1L1. Kinase activity is essential for inhibition of APC/CCDC20 and for chromosome alignment but does not play a major role in the spindle-assembly checkpoint activity. Mediates cell death in response to chromosome missegregation and acts to suppress spontaneous tumorigenesis. -
Tissue specificity
High expression in testis and thymus, less in colon, spleen, lung and small intestine. Expressed in fetal thymus, bone marrow, heart, liver, spleen and thymus. Expression is associated with cells/tissues with a high mitotic index. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. BUB1 subfamily.
Contains 1 BUB1 N-terminal domain.
Contains 1 protein kinase domain. -
Domain
The KEN box is required for its ubiquitination and degradation.
BUB1 N-terminal domain directs kinetochore localization and binding to BUB3. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Upon spindle-assembly checkpoint activation it is hyperphosphorylated and its kinase activity toward CDC20 is stimulated. Phosphorylation at Thr-609 is required for interaction with PLK1, phosphorylation at this site probably creates a binding site for the POLO-box domain of PLK1, thus enhancing the PLK1-BUB1 interaction.
Ubiquitinated and degraded during mitotic exit by APC/C-Cdh1. -
Cellular localization
Nucleus. Chromosome > centromere > kinetochore. Nuclear in interphase cells. Accumulates gradually during G1 and S phase of the cell cycle, peaks at G2/M, and drops dramatically after mitosis. Localizes to the outer kinetochore. Kinetochore localization is required for normal mitotic timing and checkpoint response to spindle damage and occurs very early in prophase. AURKB, CASC5 and INCENP are required for kinetochore localization. - Information by UniProt
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Database links
- Entrez Gene: 699 Human
- Entrez Gene: 12235 Mouse
- Entrez Gene: 296137 Rat
- Omim: 602452 Human
- SwissProt: O43683 Human
- SwissProt: O08901 Mouse
- Unigene: 469649 Human
- Unigene: 2185 Mouse
see all -
Alternative names
- Bub1 antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) antibody
see all
Images
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Western blot - Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)All lanes : Anti-Bub1 antibody [EPR18947] (ab195268) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BUB1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 122 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab195268).
Lanes 1- 2: Merged signal (red and green). Green - ab195268 observed at 125 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab195268 was shown to react with Bub1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265145 (knockout cell lysate ab257373) was used. Wild-type HeLa and BUB1 knockout HeLa cell lysates were subjected to SDS-PAGE. ab195268 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)](https://www.abcam.com/ps/products/251/ab251203/Images/ab251203-2-benefits-of-recombinant-antibodies.png)
Anti-Bub1 antibody [EPR18947] - BSA and Azide free (ab251203)
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
Certificate of Compliance
References (0)
ab251203 has not yet been referenced specifically in any publications.
