|Detection of Human Cyclin B1 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, K562 human chronic myelogenous leukemia cell line, and U2OS human osteosarcoma cell line. PVDF membrane was probed with 0.05 µg/mL of Rabbit Anti-Human Cyclin B1 Monoclonal Antibody (Catalog # MAB60001) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Cyclin B1 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Cyclin B1 in Human Squamous Cell Carcinoma. Cyclin B1 was detected in immersion fixed paraffin-embedded sections of human squamous cell carcinoma using Rabbit Anti-Human Cyclin B1 Monoclonal Antibody (Catalog # MAB60001) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.|
Intracellular Staining by Flow Cytometry
|Detection of Cyclin B1 in K562 Human Cell Line by Flow Cytometry. K562 human chronic myelogenous leukemia cell line was stained with Rabbit Anti-Human Cyclin B1 Monoclonal Antibody (Catalog # MAB60001, filled histogram) or isotype control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
Detection of Human Cyclin B1 by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Cyclin B1 at approximately 64 kDa (as indicated) using 0.5 µg/mL of Rabbit Anti-Human Cyclin B1 Monoclonal Antibody (Catalog # MAB60001). This experiment was conducted under reducing conditions and using the |
12-230 kDa separation system.
Cyclin B1 (also CCNB1 and G2/mitotic-specific cyclin-B1) is a member of the cyclin AB subfamily, cyclin family of proteins. Although its predicted MW is 50 kDa, it runs anomalously at 62 kDa in SDS-PAGE. Cyclin B1 associates with both CDK1 and 2 providing substrate specificity to a phosphorylating complex. A phosphor‑CDK1:Cyclin B1 complex is inactive and cytosolic during interphase. At the beginning of mitosis, CDK1 is dephosphorylated and activated, and the CDK1:Cyclin B1 complex initiates formation of the mitotic scaffold. Human Cyclin B1 is 433 amino acids (aa) in length. It contains two cyclin box folds (aa 201‑290 and 298‑383) and two substrate binding sites (aa 298‑342 and 343‑380). Phosphorylation occurs at Ser9, Ser35, Ser69, and Thr321. There is one potential alternative start site at Met252 and deletions of aa 363‑399 and 365‑433. Over aa 1‑91, human Cyclin B1 shares 63% aa identity with mouse Cyclin B1.
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