参阅全部 Cyclin B1 一抗
描述小鼠单克隆抗体[V152] to Cyclin B1
特异性Cyclin B1 expression is restricted to a specific short period of the cell cycle with cyclin B1 expression detected earlier and peaking in concentration before cyclin B2 expression.
经测试应用适用于: ICC/IF, IHC (PFA fixed), Flow Cyt, WB, IHC-FoFr, IHC-P, IHC-Frmore details
种属反应性与反应: Mouse, Human
Fusion protein corresponding to Hamster Cyclin B1. His-tagged Hamster Cyclin B1 expressed in bacteria, harvested from inclusion bodies, extracted with 6M guanidine HCl and purified on Nickel beads.
Database link: P14635
- WB: HeLa, Daudi, K562, Jurkat and HEK-293 whole cell lysate. IHC-P: Human tonsil tissue. Flow Cytometry: HeLa cells.
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.
存放说明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.
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
纯度Protein G purified
Our Abpromise guarantee covers the use of ab72 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC (PFA fixed)||1/400.|
|Flow Cyt||Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).
Abcam recommends using 5% Milk as the blocking agent.
|IHC-FoFr||Use a concentration of 1 - 2 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
|IHC-Fr||Use a concentration of 1 - 2 µg/ml.|
功能Essential for the control of the cell cycle at the G2/M (mitosis) transition.
序列相似性Belongs to the cyclin family. Cyclin AB subfamily.
发展阶段Accumulates steadily during G2 and is abruptly destroyed at mitosis.
翻译后修饰Ubiquitinated by the SCF(NIPA) complex during interphase, leading to its destruction. Not ubiquitinated during G2/M phases.
细胞定位Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > centrosome.
- Information by UniProt
- CCNB 1 antibody
- CCNB antibody
- ccnb1 antibody
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.
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.
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.
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.
Flow cytometry analysis of HeLa cells stained with ab72 (green line). Cell harvesting/tissue preparation method - Trypsin/EDTA. Sample Buffer: Complete Media then PBS. Fixation: Formaldehyde. Permeabilization: 90% methanol. Dilution: 1/50. Incubation time: 1 hour at 23°C, diluent: PBS/10% goat serum. Secondary antibody: Goat anti-mouse FITC at 1/1000 dilution. 100000 cells were stained, non-specific IgG antibody - negative control (purple histogram)
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.
HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained for Cyclin B1 using ab72 at a dilution of 1/500 in ICC/IF. Secondary used is a Fluorescin-conjugated Goat Anti-Mouse IgG. Primary antibody incubated for 12 hours at +4°C in TBS. Blocked using 1% BSA for 10 minutes at 25°C. Cells were permeabilized with Tween-20.
This product has been referenced in:
- Levasseur MD et al. Aneuploidy in Oocytes Is Prevented by Sustained CDK1 Activity through Degron Masking in Cyclin B1. Dev Cell 48:672-684.e5 (2019). Read more (PubMed: 30745144) »
- Daldello EM et al. Cyclin B2 is required for progression through meiosis in mouse oocytes. Development 146:N/A (2019). Read more (PubMed: 30952665) »