Anti-Cyclin B1 antibody [V152] (ab72)
![Anti-Cyclin B1 antibody [V152] (ab72)](https://www.abcam.com/ps/products/0/ab72/Images/ab72-277468-anti-cyclin-b1-antibody-v152-western-blot.jpg "Anti-Cyclin B1 antibody [V152] (ab72)")
Key features and details
- Mouse monoclonal [V152] to Cyclin B1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-Cyclin B1 antibody [V152]
See all Cyclin B1 primary antibodies -
Description
Mouse monoclonal [V152] to Cyclin B1 -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human
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Immunogen
Fusion protein within Hamster Cyclin B1. The exact sequence is proprietary.
Database link: P14635 -
Positive control
- WB: HeLa, Daudi, K562, Jurkat and HEK-293 whole cell lysate. IHC-P: Human tonsil tissue. Flow Cytometry: HeLa cells.
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General notes
This antibody clone is manufactured by Abcam.
This monoclonal antibody to cyclin B1 has been knockout validated in Western blot. The expected band for cyclin B1 was observed in wild type cells and the band was not seen in knockout cells.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
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
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
V152 -
Myeloma
Sp2 -
Isotype
IgG1 -
Light chain type
unknown -
Research areas
Images
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Western blot - Anti-Cyclin B1 antibody [V152] (ab72)
Lanes:
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Cyclin B1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab72 observed at 55 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab72 detected the expected band for Cyclin B1 in wild type HAP1 cells and the band was not seen in Cyclin B1 knockout HAP1 cells. Wild-type and Cyclin B1 knockout samples were subjected to SDS-PAGE. Ab72 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [V152] (ab72)](https://www.abcam.com/ps/products/0/ab72/Images/ab72-3288-anti-cyclin-b1-antibody-v152-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin B1 antibody [V152] (ab72)ab72 staining Cyclin B1 in human tonsil tissue - formalin-fixed paraffin-embedded section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab72, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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![Western blot - Anti-Cyclin B1 antibody [V152] (ab72)](https://www.abcam.com/ps/products/0/ab72/Images/ab72-3289-anti-cyclin-b1-antibody-v152-western-blot.jpg)
Western blot - Anti-Cyclin B1 antibody [V152] (ab72)All lanes : Anti-Cyclin B1 antibody [V152] (ab72) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
Lane 2 :Daudi whole cell lysate (ab3951)
Lane 3 : K562 (Human erythromyeloblastoid leukemia cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) whole cell lysate
Lane 5 : HEK-293 (Human embryonic kidney cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 48 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa, 77 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
Abcam recommends using milk as the blocking agent. -
![Western blot - Anti-Cyclin B1 antibody [V152] (ab72)](https://www.abcam.com/ps/products/0/ab72/Images/ab72-3287-anti-cyclin-b1-antibody-v152-western-blot.jpg)
Western blot - Anti-Cyclin B1 antibody [V152] (ab72)Lanes 1-9 shows the Cyclin B1 wave as HeLa cells enter and exit mitosis. HeLa cells were synchronised in the early S phase by double thymidine block then released to synchronously enter mitosis. Time points were taken at the points indicated.
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate loaded at 25 µg per lane. ab72 used at 1/500 in reducing conditions in conjunction with goat anti mouse (HRP) 1/20,000.
This image is courtesy of an Abreview submitted on 12 September 2005. We do not have any further information relating to this image.
