WB result of Cyclin B1 Recombinant Rabbit mAb
Primary antibody: Cyclin B1 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HEK-293 whole cell lysate 20 µg
Lane 3: U-2 OS whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 48 kDa
Observed MW: 55 kDa
Cyclin B1 Recombinant Rabbit mAb (S-1407-224)
Cyclin B1 Recombinant Rabbit mAb (S-1407-224)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Cyclin B1 |
| Synonyms | G2/mitotic-specific cyclin-B1; CCNB1; CCNB |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | P14635 |
| Clone Number | S-1407-224 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu |
| Positive Sample | HeLa, HEK-293, U-2 OS |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu |
| IHC-P | 1:250 | Hu |
| ICC | 1:500 | Hu |
| ICFCM | 1:50 | Hu |
Background
Cyclin B1 is a crucial regulatory protein involved in the cell cycle, particularly in controlling the G2/M transition phase. It forms transiently active kinase complexes with CDK1 (Cyclin-dependent kinase 1) that play distinct roles in mitosis. It accumulates during S-G2, shuttles between the cytoplasm and nucleus, and becomes concentrated in the nucleus in late G2. During prometaphase, Cyclin B1 localizes to centrosomes, mitotic spindle, and chromosomes. Cyclin B1 is involved in the attachment of kinetochore fibers (k-fibers) formed by microtubules during mitosis. It is degraded after the completion of microtubule attachment to kinetochores during mitosis and inactivation of the spindle assembly checkpoint. Uncontrolled expression of Cyclin B1 is associated with neoplastic transformation and the development of gynecological cancers. It has been implicated in the progression of various types of cancer, including gastric, breast, and non-small cell lung cancer. Elevated Cyclin B1 expression correlates with decreased overall survival in multiple cancer types. Cyclin B1/CDK1 is also involved in mitochondrial bioenergetics, which plays a pivotal role in tumor radiation response and resistance to anti-cancer therapies.
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) labelling Cyclin B1 antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human placenta. Anti-Cyclin B1 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-Cyclin B1 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human tonsil. Anti-Cyclin B1 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Cyclin B1 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
