Anti-CENPB antibody (ab25734)
Key features and details
- Rabbit polyclonal to CENPB
- Suitable for: WB, ICC/IF
- 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 & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human CENPB.
Read Abcam's proprietary immunogen policy (Peptide available as ab28019.) -
Positive control
- ICC: HeLa cells. WB: A431, Jurkat, Daudi and HeLa cell lysates.
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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications MouseRatSheepHamsterApplication Abreviews Notes WB (1) Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).ICC/IF (4) 1/200 - 1/1000.Notes 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.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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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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Western blot - Anti-CENPB antibody (ab25734)All lanes : Anti-CENPB antibody (ab25734) at 1 µg/ml
Lane 1 : Wild-type A431 cell lysate
Lane 2 : CENPB knockout A431 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Daudi 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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Western blot - Anti-CENPB antibody (ab25734)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/ Immunofluorescence - 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. -
Immunocytochemistry/ Immunofluorescence - Anti-CENPB antibody (ab25734)ICC/IF image of ab25734 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab25734, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HEK 293 cells.
Protocols
References (33)
ab25734 has been referenced in 33 publications.
- Barra V et al. Phosphorylation of CENP-A on serine 7 does not control centromere function. Nat Commun 10:175 (2019). PubMed: 30635586
- Cheng Z et al. circTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1. Nat Commun 10:3200 (2019). PubMed: 31324812
- Addis Jones O et al. PLK1 facilitates chromosome biorientation by suppressing centromere disintegration driven by BLM-mediated unwinding and spindle pulling. Nat Commun 10:2861 (2019). PubMed: 31253795
- Choi SH et al. CDK12 phosphorylates 4E-BP1 to enable mTORC1-dependent translation and mitotic genome stability. Genes Dev 33:418-435 (2019). PubMed: 30819820
- Kyriacou E & Heun P High-resolution mapping of centromeric protein association using APEX-chromatin fibers. Epigenetics Chromatin 11:68 (2018). PubMed: 30445992
Images
-
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).
-
Western blot - Anti-CENPB antibody (ab25734)All lanes : Anti-CENPB antibody (ab25734) at 1 µg/ml
Lane 1 : Wild-type A431 cell lysate
Lane 2 : CENPB knockout A431 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Daudi 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.
-
Western blot - Anti-CENPB antibody (ab25734)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/ Immunofluorescence - 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.
-
Immunocytochemistry/ Immunofluorescence - Anti-CENPB antibody (ab25734)
ICC/IF image of ab25734 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab25734, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HEK 293 cells.
