AB209998

Anti-BubR1 antibody [EPR20652]

RabMAb
Recombinant
What is this?

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(2 Publications)

Rabbit Recombinant Monoclonal BubR1 antibody. Suitable for IP, WB, ICC/IF and reacts with Human samples. Cited in 2 publications.

View Alternative Names

BUBR1, MAD3L, SSK1, BUB1B, Mitotic checkpoint serine/threonine-protein kinase BUB1 beta, MAD3/BUB1-related protein kinase, Mitotic checkpoint kinase MAD3L, Protein SSK1, hBUBR1

7 Images

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of A549 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HT-29 (human colorectal adenocarcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HT-29 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling BubR1 with ab209998 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling BubR1 with ab209998 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing specific staining on centromeres of HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution.
-ve control : Secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

Lab

Immunoprecipitation - Anti-BubR1 antibody [EPR20652] (AB209998)

BubR1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209998 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209998 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).

Lane 2 : ab209998 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab209998 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-BubR1 antibody [EPR20652] (ab209998) at 1/1000 dilution

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>)

Predicted band size: 119 kDa

Observed band size: 120 kDa

false

Exposure time: 1s

Lab

Western blot - Anti-BubR1 antibody [EPR20652] (AB209998)

Positive BubR1 detection by WB is dependent on the number of mitotic cells in the sample and may need optimization for some cell and tissue lysates.

Blocking/Dilution buffer : 5% NFDM/TBST

Exposure time : Lane 1 - 10 seconds, lane 2 - 3 seconds, lane 3 - 3 minutes

All lanes:

Western blot - Anti-BubR1 antibody [EPR20652] (ab209998) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

Raji (human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

Lane 3:

HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 119 kDa

Observed band size: 120 kDa

false

Carrier free

Anti-BubR1 antibody [EPR20652] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR20652

Isotype

IgG

Carrier free

Reacts with

Human

Applications

ICC/IF, WB, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "" } } }

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Product details

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

BubR1 also known as BUB1B is an important protein in the mitotic checkpoint complex with an approximate mass of 120 kDa. BubR1 ensures proper chromosome alignment and segregation during cell division by inhibiting the anaphase-promoting complex/cyclosome (APC/C) until all chromosomes achieve proper attachment to the spindle. This protein localizes mainly to the kinetochores of chromosomes and is commonly expressed in dividing cells throughout the body.

Biological function summary

The spindle assembly checkpoint mechanism involves a complex in which BubR1 plays a central role. BubR1 along with other checkpoint proteins such as Bub1 and Mad2 forms a surveillance complex ensuring the correct attachment of microtubules to kinetochores. This action prevents premature progression to anaphase reducing the risk of aneuploidy and maintaining genomic stability.

Pathways

Mitotic checkpoint control critically involves BubR1 linking closely with several proteins including Mad2 and Cdc20. BubR1 integrates into the spindle checkpoint pathway a central player in regulating mitosis. Within this pathway BubR1 inhibits APC/CCdc20 activity preventing premature anaphase onset until all chromosomes are correctly oriented to divide. Interactions between these proteins ensure accurate mitotic progression and cellular replication fidelity.

BubR1 function directly impacts cancer development and premature aging syndromes. Defective BubR1 activity can lead to chromosomal instability a characteristic of many cancers including colorectal cancer. Moreover reduced BubR1 expression links with premature aging syndromes such as mosaic variegated aneuploidy through its interaction with proteins such as p53 highlighting its role in genome integrity maintenance and cellular aging processes.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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.

See full target information BUB1B

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Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Communications biology 6:249 PubMed36882565

2023

Ribonucleotide reductase subunit switching in hepatoblastoma drug response and relapse.

Applications

Unspecified application

Species

Unspecified reactive species

Anthony Brown,Qingfei Pan,Li Fan,Emilie Indersie,Cheng Tian,Nikolai Timchenko,Liyuan Li,Baranda S Hansen,Haiyan Tan,Meifen Lu,Junmin Peng,Shondra M Pruett-Miller,Jiyang Yu,Stefano Cairo,Liqin Zhu

Nature aging 3:185-201 PubMed37118121

2023

Unique progerin C-terminal peptide ameliorates Hutchinson-Gilford progeria syndrome phenotype by rescuing BUBR1.

Applications

Unspecified application

Species

Unspecified reactive species

Na Zhang,Qianying Hu,Tingting Sui,Lu Fu,Xinglin Zhang,Yu Wang,Xiaojuan Zhu,Baiqu Huang,Jun Lu,Zhanjun Li,Yu Zhang

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-BubR1 antibody [EPR20652]

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunocytochemistry/ Immunofluorescence - Anti-BubR1 antibody [EPR20652] (AB209998)

Immunoprecipitation - Anti-BubR1 antibody [EPR20652] (AB209998)

Western blot - Anti-BubR1 antibody [EPR20652] (AB209998)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (2)

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