Anti-CENPB antibody (ab25734)
Key features and details
- Rabbit polyclonal to CENPB
- Suitable for: WB, ICC/IF, ICC
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-CENPB antibody
See all CENPB primary antibodies -
Description
Rabbit polyclonal to CENPB -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, ICCmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Sheep, Hamster -
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human CENPB.
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Positive control
- ICC: HeLa cells. WB: A431, Jurkat, Daudi and HeLa cell lysates.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab25734 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 |
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WB | (1) |
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
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ICC/IF | (5) |
1/200 - 1/1000.
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ICC |
Use a concentration of 1 µg/ml.
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Notes |
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WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa). |
ICC/IF
1/200 - 1/1000. |
ICC
Use a concentration of 1 µg/ml. |
Target
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Function
Interacts with centromeric heterochromatin in chromosomes and binds to a specific subset of alphoid satellite DNA, called the CENP-B box. May organize arrays of centromere satellite DNA into a higher order structure which then directs centromere formation and kinetochore assembly in mammalian chromosomes. -
Sequence similarities
Contains 1 HTH CENPB-type DNA-binding domain.
Contains 1 HTH psq-type DNA-binding domain. -
Cellular localization
Nucleus. Chromosome > centromere. - Information by UniProt
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Database links
- Entrez Gene: 1059 Human
- Entrez Gene: 12616 Mouse
- Entrez Gene: 362217 Rat
- Omim: 117140 Human
- SwissProt: P07199 Human
- SwissProt: P27790 Mouse
- SwissProt: P49451 Sheep
- Unigene: 516855 Human
see all -
Alternative names
- CENP B antibody
- CENP-B antibody
- CENPB antibody
see all
Images
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All lanes : Anti-CENPB antibody (ab25734) at 1 µg/ml
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : CENPB knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 77 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab25734 observed at 77 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab25734 was shown to react with CENPB in wild-type A431 cells in Western blot with loss of signal observed in CENPB knockout sample.Wild-type A431 and CENPB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab25734 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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ab25734 staining CENPB in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab25734 at 1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in green) and ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Immunocytochemistry/ Immunofluorescence - Anti-CENPB antibody (ab25734)Image courtesy of an AbReview by Weiguo Zhang
4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for CENPB (red) using ab25734 at 1/200 dilution in ICC/IF.
The nuclear counterstain is DAPI (blue). Alpha-Tubulin is stained with an mouse monoclonal anti-alpha Tubulin antibody (green).
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All lanes : Anti-CENPB antibody (ab25734) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A-431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 65 kDa
Observed band size: 65 kDa
Additional bands at: 74 kDa (possible cross reactivity), 75 kDa (possible cross reactivity) -
Immunocytochemistry - Anti-CENPB antibody (ab25734)This image is courtesy of an Abreview submitted by Dr Kirk McManusab25734 (1/1000) staining CENPB in HeLa cells (green). Cells were fixed with methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (44)
ab25734 has been referenced in 44 publications.
- Tian W et al. LINC01123 potentially correlates with radioresistance in glioma through the miR-151a/CENPB axis. Neuropathology 42:3-15 (2022). PubMed: 34519373
- Chen Y et al. Centromeric transcription maintains centromeric cohesion in human cells. J Cell Biol 220:N/A (2021). PubMed: 33881484
- Hou H et al. Centromeres are dismantled by foundational meiotic proteins Spo11 and Rec8. Nature 591:671-676 (2021). PubMed: 33658710
- Murillo-Pineda M et al. Induction of spontaneous human neocentromere formation and long-term maturation. J Cell Biol 220:N/A (2021). PubMed: 33443568
- Dumont M et al. Human chromosome-specific aneuploidy is influenced by DNA-dependent centromeric features. EMBO J 39:e102924 (2020). PubMed: 31750958