Human Cyclin B1 Antibody

Recombinant Monoclonal Antibody.
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Cyclin B1 in direct ELISAs and Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 2061D
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E. coli-derived recombinant human Cyclin B1
    Met1-Pro91
    Accession # P14635
  • Formulation
    Supplied as a solution in PBS containing BSA, Glycerol and Sodium Azide. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.05 µg/mL
    See below
  • Simple Western
    0.5 µg/mL
    See below
  • Immunohistochemistry
    3-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunohistochemistry
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.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 °C, as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
Background: Cyclin B1

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.

  • Entrez Gene IDs:
    891 (Human); 268697 (Mouse); 25203 (Rat)
  • Alternate Names:
    CCNB; CCNB1; Cyclin B1; G2/mitotic-specific cyclin B1; G2/mitotic-specific cyclin-B1
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Isotype Controls
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Normal Rabbit IgG Control

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Normal Rabbit IgG Control

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Secondary Antibodies
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Rabbit IgG VisUCyte HRP Polymer Antibody

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Donkey Anti-Rabbit IgG (H+L) Antibody

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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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 WB Human Anonymous 08/16/2017
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Summary

ApplicationWestern Blot
Sample TestedT47D human breast cancer cell line,MCF-7 human breast cancer cell line
SpeciesHuman

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