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Immunofluorescent analysis of Cyclin B1 (green) showing staining in the in the cytoplasm and nucleus of C2C12 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cyclin B1 monoclonal antibody (Product # MA1-155) in 3% BSA-PBS at a dilution of 1:150 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Hamster, Human, Mouse, Primate, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Hamster Cyclin B1|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10 - 1:2000|
|Immunoprecipitation (IP)||1 µg|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-155 detects Cyclin B1 in mammalian samples. MA1-155 has been successfully used in Immunohistochemistry (paraffin), Immunofluorescence, Immunoprecipitation, Flow Cytometry and Western Blot procedures. Immunoprecipitation and Western Blot analysis with MA1-155 show the accumulation of a prominent band at ~52 kDa in camptothecin and hydroxyurea treated cells. MA1-155 also detects additional unknown bands at ~35 and ~75 kDa. In Immunofluorescence applications, MA1-155 shows cyclin B1 co-localization with microtubules, whereas MA1-156 shows cyclin B2 staining consistent with the Golgi region.
MA1-155 specifically detects Cyclin B1 in in whole cell lysates, but will also cross-react with purified recombinant human Cyclin B2 in Western Blot applications.
Cyclins bind to and regulate the activity of the Cyclin Dependent Protein Kinases (CDKs). The protein encoded by the CCNB1 gene is a regulatory protein involved in mitosis. The gene product complexes with p34(cdc2) to form the maturation-promoting factor (MPF). Two alternative transcripts have been found, a constitutively expressed transcript and a cell cycle-regulated transcript, that is expressed predominantly during G2/M phase. Cyclin B1 is not ubiquitinated during G2/M phase, resulting in its steady accumulation during G2 phase, followed by abrupt APC dependent destruction at the end of mitosis. Destruction of Cyclin B1 is required for cell cycle progression. Cyclin B1 is overexpressed in various cancers, including breast, prostate, and non-small cell lung cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
CCNB; CCNB1; cyclin B1; cyclin B1, related sequence 1; cyclin B1, related sequence 13; G2/Mitotic-Specific Cyclin B1; G2/mitotic-specific cyclin-B1
Ccn-2; CCNB; CCNB1; Ccnb1-rs1; Ccnb1-rs13; Cycb; Cycb-4; Cycb-5; CycB1; Cycb1-rs1