Recombinant Anti-BubR1 antibody [EPR22544-48] (ab254326)

Product name

Anti-BubR1 antibody [EPR22544-48]
See all BubR1 primary antibodies
Description

Rabbit monoclonal [EPR22544-48] to BubR1
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt,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: HeLa and Raji cell lysate; Mouse testis, thymus and rat testis tissue lysate. IHC-P: Human breast cancer and testis tissue; Mouse and rat testis tissue. ICC/IF: SK-OV-3 and HeLa cells.
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: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22544-48
Isotype

IgG
Research areas

Alternative Versions
- Anti-BubR1 antibody [EPR22544-48] - BSA and Azide free (ab255249)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Recombinant Protein
- Recombinant human BubR1 protein (ab133149)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab254326 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
WB		1/1000. Predicted molecular weight: 119 kDa.
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/50.

Notes
WB 1/1000. Predicted molecular weight: 119 kDa.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.

Application notes

Is unsuitable for Flow Cyt,IHC-Fr or IP.

Function

Essential component of the mitotic checkpoint. Required for normal mitosis progression. The mitotic checkpoint delays anaphase until all chromosomes are properly attached to the mitotic spindle. One of its checkpoint functions is to inhibit the activity of the anaphase-promoting complex/cyclosome (APC/C) by blocking the binding of CDC20 to APC/C, independently of its kinase activity. The other is to monitor kinetochore activities that depend on the kinetochore motor CENPE. Required for kinetochore localization of CENPE. Negatively regulates PLK1 activity in interphase cells and suppresses centrosome amplification. Also implicated in triggering apoptosis in polyploid cells that exit aberrantly from mitotic arrest. May play a role for tumor suppression.
Tissue specificity

Highly expressed in thymus followed by spleen. Preferentially expressed in tissues with a high mitotic index.
Involvement in disease

Note=Defects in BUB1B are associated with tumor formation.
Defects in BUB1B are the cause of premature chromatid separation trait (PCS) [MIM:176430]. PCS consists of separate and splayed chromatids with discernible centromeres and involves all or most chromosomes of a metaphase. It is found in up to 2% of metaphases in cultured lymphocytes from approximately 40% of normal individuals. When PCS is present in 5% or more of cells, it is known as the heterozygous PCS trait and has no obvious phenotypic effect, although some have reported decreased fertility. Inheritance is autosomal dominant.
Defects in BUB1B are the cause of mosaic variegated aneuploidy syndrome (MVA) [MIM:257300]. MVA is a severe autosomal recessive developmental disorder characterized by mosaic aneuploidies, predominantly trisomies and monosomies, involving multiple different chromosomes and tissues. The proportion of aneuploid cells varies but is usually more than 25% and is substantially greater than in normal individuals. Affected individuals typically present with severe intrauterine growth retardation and microcephaly. Eye anomalies, mild dysmorphism, variable developmental delay, and a broad spectrum of additional congenital abnormalities and medical conditions may also occur. The risk of malignancy is high, with rhabdomyosarcoma, Wilms tumor and leukemia reported in several cases. MVA is caused by biallelic mutations in the BUB1B gene.
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 D-box targets the protein for rapid degradation by ubiquitin-dependent proteolysis during the transition from mitosis to interphase.
The BUB1 N-terminal domain directs kinetochore localization and binding to BUB3.
Post-translational
modifications

Proteolytically cleaved by caspase-3 in a cell cycle specific manner. The cleavage might be involved in the durability of the cell cycle delay. Caspase-3 cleavage is associated with abrogation of the mitotic checkpoint. The major site of cleavage is at Asp-610.
Acetylation at Lys-250 regulates its degradation and timing in anaphase entry.
Ubiquitinated. Degradated by the proteasome.
Sumoylated by SUMO2 and SUMO3. The sumoylation mediates the association with CENPE at the kinetochore.
Autophosphorylated in vitro. Intramolecular autophosphorylation is stimulated by CENPE. Phosphorylated during mitosis and hyperphosphorylated in mitotically arrested cells. Phosphorylation at Ser-670 and Ser-1043 occurs at kinetochores upon mitotic entry with dephosphorylation at the onset of anaphase.
Cellular localization

Cytoplasm. Nucleus. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > centrosome. Cytoplasmic in interphase cells. Associates with the kinetochores in early prophase. Kinetochore localization requires BUB1, PLK1 and CASC5.
Target information above from: UniProt accession O60566 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 701 Human
- Entrez Gene: 12236 Mouse
- Entrez Gene: 171576 Rat
- Omim: 602860 Human
- SwissProt: O60566 Human
- SwissProt: Q9Z1S0 Mouse
- Unigene: 513645 Human
- Unigene: 29133 Mouse
Alternative names
- Beta homolg of S. cerevisiae BUB 1 antibody
- Beta homolg of S. cerevisiae budding uninhibited by benzimidazoles antibody
- BUB 1B antibody
- BUB1 budding uninhibited by benzimidazoles 1 homolog beta antibody
- Bub1A antibody
- BUB1B antibody
- BUB1B_HUMAN antibody
- BUB1beta antibody
- BUBR1 antibody
- Budding Uninhibited by Benzimidazoles 1 beta antibody
- Budding uninhibited by benzimidazoles 1 homolog beta (yeast) antibody
- hBUBR1 antibody
- MAD3/BUB1 related protein kinase antibody
- MAD3/BUB1-related protein kinase antibody
- MAD3L antibody
- Mitotic checkpoint gene BUB1B antibody
- Mitotic checkpoint kinase MAD3L antibody
- Mitotic checkpoint serine/threonine protein kinase BUB1 beta antibody
- Mitotic checkpoint serine/threonine-protein kinase BUB1 beta antibody
- MVA1 antibody
- OTTHUMP00000160319 antibody
- Protein SSK1 antibody
- SSK 1 antibody
- SSK1 antibody
see all

Western blot - Anti-BubR1 antibody [EPR22544-48] (ab254326)

All lanes : Anti-BubR1 antibody [EPR22544-48] (ab254326) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse thymus tissue lysate
Lane 5 : Rat testis tissue 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

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

The molecular weight observed is consistent with what has been described in the literature (PMID: 26889953).

Exposure time:

Lanes 1-2: 70 seconds

Lane 3: 48 seconds

Lanes 4-5: 3 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling BubR1 with ab254326 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat testis (PMID: 20411023). The section was incubated with ab254326 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling BubR1 with ab254326 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse testis (PMID: 20411023). The section was incubated with ab254326 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling BubR1 with ab254326 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human testis (PMID: 20411023). The section was incubated with ab254326 for 30 mins at room temperature. Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling BubR1 with ab254326 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human breast cancer is observed. The section was incubated with ab254326 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (human ovarian cancer epithelial cell) cells labeling BubR1 with ab254326 at 1/50 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing specific staining on centromeres of SK-OV-3 cell line in M phase. The nuclear counterstain is DAPI (Blue). Tubulin was stained using Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution
Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling BubR1 with ab254326 at 1/50 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green).Confocal image showing cytoplasmic staining in HeLa cell line. The nuclear counterstain is DAPI (Blue). Tubulin was stained using Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution
Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR22544-48] (ab254326)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling BubR1 with ab254326 at 1/50 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing specific staining on centromeres of HeLa cell line in M phase. The nuclear counterstain is DAPI (Blue). Tubulin was stained using Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution
Anti-BubR1 antibody [EPR22544-48] (ab254326)

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

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Datasheet download

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Publishing research using ab254326? Please let us know so that we can cite the reference in this datasheet.

ab254326 has been referenced in 2 publications.

Ding ZM et al. Diethylstilbestrol exposure disrupts mouse oocyte meiotic maturation in vitro through affecting spindle assembly and chromosome alignment. Chemosphere 249:126182 (2020). PubMed: 32078850
Zhang SX et al. Bisphenol B Exposure Disrupts Mouse Oocyte Meiotic Maturation in vitro Through Affecting Spindle Assembly and Chromosome Alignment. Front Cell Dev Biol 8:616771 (2020). PubMed: 33392205

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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

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications