Cyclin B1 antibody

Catalog No. ABIN967429

Product Details anti-Cyclin B1 Antibody

Target: Cyclin B1 (CCNB1)

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This Cyclin B1 antibody is un-conjugated

Application: Western Blotting (WB), Flow Cytometry (FACS), Immunoprecipitation (IP), BioImaging (BI)

Brand: BD Pharmingen™

Cross-Reactivity: Mouse (Murine)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Recombinant Human Cyclin B1

Clone: GNS-11

Isotype: IgG2a

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methnaol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389, ABIN968537

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C.

Target Details for Cyclin B1

Target: Cyclin B1 (CCNB1)

Alternative Name: Cyclin B1 (CCNB1 Products)

Synonyms: ccnb antibody, cycb antibody, cb267 antibody, cycb1 antibody, wu:fa19g04 antibody, wu:fb16d01 antibody, wu:fb16e07 antibody, wu:fi21c01 antibody, ccnb1 antibody, MGC53596 antibody, CCNB antibody, Ccnb1-rs1 antibody, Ccnb1-rs13 antibody, CycB1 antibody, Cycb-4 antibody, Cycb-5 antibody, Cycb1-rs1 antibody, cyclin B1 antibody, cyclin B1 S homeolog antibody, ccnb1 antibody, ccnb1.2 antibody, ccnb1.S antibody, CCNB1 antibody, Ccnb1 antibody, ccnb1.2.S antibody

Background: During the cell cycle, most eukaryotic cells double in mass, replicate their DNA and then distribute identical copies of their genome to progeny cells during mitosis. An internal biochemical clock ensures that all the necessary events occur in proper sequence and at the appropriate time. Cell in M phase contain a dominant regulatory factor known as maturation promoting factor (MPF). MPF triggers a variety of enzymatic and ultrastructural changes that are necessary for cell division. In higher eukaryotes, these mitosis-specific alterations include disassembly of the nuclear envelope, packaging of the DNA into chromosomes and assembly of the mitotic spindle. Purified MPF is a regulator of a protein kinase cascade and is evolutionarily conserved in all eukaryotic cells ranging from yeast to man. MPF consists predominantly of two polypeptides, cyclin B1 and p34, and contains protein kinase activity itself. It is the major M-phase-specific histone H1 kinase, but also phosphorylates a variety of other substrates including lamins, nucleolin, RNA polymerase II, retinoblastoma protein, SV40 large T antigen, p53, and the oncogenes c-src and c-abl. Cyclin B1 migrates at a reduced molecular weight of 62 kDa on SDS-PAGE.

Molecular Weight: 62 kDa

Pathways: Cell Division Cycle, AMPK Signaling, Mitotic G1-G1/S Phases, M Phase