Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

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Anti-BubR1 antibody [EPR20652] (ab209998)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20652] to BubR1
Suitable for: WB, ICC/IF, IP
Reacts with: Human

Product name

Anti-BubR1 antibody [EPR20652]
See all BubR1 primary antibodies
Description

Rabbit monoclonal [EPR20652] to BubR1
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human cell lines HeLa, Raji and HepG2 whole cell lysates. ICC/IF: Human cell lines HeLa, HepG2, A549 and HT-29. IP: Human cell line HeLa whole cell lysate.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20652
Isotype

IgG
Research areas

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

All lanes :

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366)

Observed band size: 120 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second
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.
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.
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 subcellular localization of BubR1 changes through mitosis of HeLa cell line (PMID: 25833949). The nuclear counter stain is DAPI (blue).
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.
Western blot - Anti-BubR1 antibody [EPR20652] (ab209998)

All lanes : Anti-BubR1 antibody [EPR20652] (ab209998) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Performed under reducing conditions.

Predicted band size: 119 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?

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
Anti-BubR1 antibody [EPR20652] (ab209998)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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