Invitrogen

Cyclin B1 Monoclonal Antibody (GNS11)

Advanced Verification

This Antibody was verified by Cell Treatment to ensure that the antibody binds to the antigen stated. View Details

9 References

View all (353) Cyclin B1 antibodies

Datasheet

Datasheet

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Cyclin B1 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on formalin-fixed, paraffin-embedded human tonsil tissue. To expose target proteins, heat-induced antigen retrieval was performed by boiling tissue sections in 10mM sodium citrate buffer (pH 6) for 10-20 minutes. Tissues were probed with a Cyclin B1 monoclonal antibody (Product # MA5-14327) at a concentration of 3 µg/mL. Detection was performed using an anti-mouse IgG-HRP secondary antibody followed by a colorimetric substrate.

Cyclin B1 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of Cyclin B1 was performed on the Human Colon Carcinoma cell line LS174T. Antigen-antibody complexes were formed by incubating 500 µL of LS174T whole cell lysate with 5 µL of a Cyclin B1 monoclonal antibody (Product # MA5-14327). The immune complexes were captured using Protein A agarose, washed extensively, and eluted. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a membrane, and probed with a series of reagents to detect Cyclin B1.

Cyclin B1 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Cyclin B1 (green) in HeLa cells either left untreated (left panel) or treated with 50nM Nocodazole (right panel) for 16 hours. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Cyclin B1 monoclonal antibody (Product # MA5-14327) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.

Cyclin B1 Antibody in Western Blot (WB) — Western blot analysis of Cyclin B1 was performed by loading 25 µg of HT29 (Lane 1) and Jurkat (Lane 2)cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Cyclin B1 monoclonal antibody (Product # MA5-14327) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at approx. 58 kDa.

Cyclin B1 Antibody in Cell Treatment — Modulation of expression of target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis of Cyclin B1 using Cyclin B1 monoclonal antibody (Product # MA5-14327) shows induced expression of Cyclin B1 in the cytoplasm of HeLa cells treated with Nocodazole. {TM}

Product Details

Tested Applications

Dilution

Immunocytochemistry (ICC)

1:10-1:200

Immunofluorescence (IF)

1:10-1:200

Immunohistochemistry (Paraffin) (IHC (P))

2-4 µg/mL

Immunoprecipitation (IP)

2 µg/mL

Western Blot (WB)

1:100-1:1000

Published Applications

Western Blot (WB)

See 3 publications below

Immunocytochemistry (ICC)

See 2 publications below

Immunohistochemistry (IHC)

See 3 publications below

Immunoprecipitation (IP)

See 1 publication below

Product Specifications

Species Reactivity

Hamster, Human, Mouse, Rat

Published species

Human, Mouse, Rat

Host / Isotype

Mouse / IgG2a

Class

Monoclonal

Type

Antibody

Clone

GNS11

Immunogen

Human cyclin B1 protein

Conjugate

Unconjugated

Form

Liquid

Concentration

0.2 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10980440

Product Specific Information

MA5-14327 targets Cyclin B1 in WB, ICC/IF, IHC (P), and IP applications and shows reactivity with Hamster, Human, mouse, and Rat samples. This antibody is not recommended for NIH-3T3 cells in Western blot applications.

The MA5-14327 immunogen is human cyclin B1 protein.

This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.

Target Information

Cyclins bind to and regulate the activity of the Cyclin dependent protein kinases (CDKs). Cyclin B1 or CCNB1 is a regulatory protein involved in mitosis. It is essential for the control of the cell cycle at the G2/M transition. Cyclin B1 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.

Bioinformatics

Protein Aliases: cyclin B1; cyclin B1, related sequence 1; cyclin B1, related sequence 13; G2/mitotic-specific cyclin B1; G2/mitotic-specific cyclin-B1; OTTHUMP00000223023

Gene Aliases: Ccn-2; CCNB; CCNB1; Ccnb1-rs1; Ccnb1-rs13; Cycb; Cycb-4; Cycb-5; CycB1; Cycb1-rs1; I79_013589

UniProt ID: (Human) P14635, (Chinese hamster) Q08301, (Rat) P30277, (Mouse) P24860

Entrez Gene ID: (Human) 891, (Chinese hamster) 100770842, (Rat) 25203, (Mouse) 268697

Molecular Function: enzyme modulator kinase activator kinase modulator