Cyclin B1 (CCNB1) (AA 1-21) antibody

Details for Product No. ABIN967427
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Antigen
Synonyms CCNB, Ccnb1-rs1, Ccnb1-rs13, CycB1, Cycb-4, Cycb-5, Cycb1-rs1, ccnb, ccnb1, cycb, cb267, cycb1, wu:fa19g04, wu:fb16d01, wu:fb16e07, wu:fi21c01, MGC53596
Epitope
AA 1-21
(34), (27), (19), (13), (5), (4), (4), (4), (3), (3), (3), (3), (3), (3), (3), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(244), (95), (88), (27), (26), (16), (13), (13), (12), (2), (1), (1), (1)
Host
Mouse
(202), (62)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(7), (4), (4), (4), (4), (4), (4), (4), (4), (4), (4)
Application
Western Blotting (WB), BioImaging (BI), Flow Cytometry (FACS), Fluorescence Microscopy (FM), Immunohistochemistry (IHC), Immunoprecipitation (IP)
(177), (78), (71), (69), (53), (43), (40), (18), (14), (7), (3), (2), (1), (1)
Pubmed 13 references available
Catalog no. ABIN967427
Quantity 0.25 mg
Price
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Immunogen Human Cyclin B1 Recombinant Protein
Clone GNS-1
Isotype IgG1
Cross-Reactivity Hamster, 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 Purified from tissue culture supernatant or ascites by affinity chromatography.
Purity Purified
Alternative Name Cyclin B1
Background Cyclins and cyclin-dependent kinases (cdks) are evolutionarily conserved proteins that are essential for cell-cycle control in eukaryotes. Cyclins (regulatory subunits) bind to cdks (catlytic subunits) to form complexes that regulate the progression of the cell cycle. The main cyclin-cdks complexes formed in vertebrate cells are cyclin D-cdk4 (G0/G1), cyclin E-cdk2 (G1/S), cyclin A-cdk2 (S) and cyclin B1-cdk1 (G2/M). These complexes are regulated by activating and inhibitory phosphorylation events, as well as by interactions with small regulatory proteins, such as p21 and p27 [Kip1]. Cyclin B1 is a mitotic cyclin, where expression is normally low in G0/G1, increases in S and is maximal during the G2/M phase. Cyclin B1 is rapidly degraded at the end of mitosis, and is required for cells to exit from mitosis. This antibody has been reported to react to hamster and mouse cyclin B1. In addition, the GNS-1 antibody has been reported to recognize an epitope between amino acids 1-21 of human cyclin B1.
Molecular Weight 62 kDa
Research Area Cancer, Cell Cycle
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% methanol, 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 min 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 hr 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 dark for 1 hr 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 min before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
Comment

Related Products: ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 0.5 mg/ml
Buffer Aqueous buffered solution.
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
Supplier Images
anti-Cyclin B1 (CCNB1) (AA 1-21) antibody Western blot analysis of cyclin B1. Lane 1: K562 human leukemia cell lysate. Lane 2: 293 human embryonic kidney cell lysate. Anti-human cyclin B1 (ABIN967426) identifies cyclin B1 as an ~62 kDa band.
anti-Cyclin B1 (CCNB1) (AA 1-21) antibody (2) Cyclin B1 staining of U-2 OS (ATCC HTB-96) cells. Cells were seeded in a 96 well imaging plate at ~10,000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-cyclin B1 antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen). Images were taken on a BD Pathway™ 855 Bioimager system using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and HeLa (CCL-2) cells and worked with both the Triton™ X-100 and alcohol perm protocols.
General Cao, Faha, Dembski et al.: "Independent binding of the retinoblastoma protein and p107 to the transcription factor E2F." in: Nature, Vol. 355, Issue 6356, pp. 176-9, 1992 (PubMed).

Faha, Ewen, Tsai et al.: "Interaction between human cyclin A and adenovirus E1A-associated p107 protein." in: Science (New York, N.Y.), Vol. 255, Issue 5040, pp. 87-90, 1992 (PubMed).

Gong, Traganos, Darzynkiewicz: "Discrimination of G2 and mitotic cells by flow cytometry based on different expression of cyclins A and B1." in: Experimental cell research, Vol. 220, Issue 1, pp. 226-31, 1995 (PubMed).

Coleman, Tang, Dunphy: "Negative regulation of the wee1 protein kinase by direct action of the nim1/cdr1 mitotic inducer." in: Cell, Vol. 72, Issue 6, pp. 919-29, 1993 (PubMed).

Zhang, Xiong, Beach: "Proliferating cell nuclear antigen and p21 are components of multiple cell cycle kinase complexes." in: Molecular biology of the cell, Vol. 4, Issue 9, pp. 897-906, 1994 (PubMed).

Gong, Ardelt, Traganos et al.: "Unscheduled expression of cyclin B1 and cyclin E in several leukemic and solid tumor cell lines." in: Cancer research, Vol. 54, Issue 16, pp. 4285-8, 1994 (PubMed).

Sherwood, Rush, Kung et al.: "Cyclin B1 expression in HeLa S3 cells studied by flow cytometry." in: Experimental cell research, Vol. 211, Issue 2, pp. 275-81, 1994 (PubMed).

Gong, Traganos, Darzynkiewicz: "Simultaneous analysis of cell cycle kinetics at two different DNA ploidy levels based on DNA content and cyclin B measurements." in: Cancer research, Vol. 53, Issue 21, pp. 5096-9, 1993 (PubMed).

Kung, Sherwood, Schimke: "Differences in the regulation of protein synthesis, cyclin B accumulation, and cellular growth in response to the inhibition of DNA synthesis in Chinese hamster ovary and HeLa S3 cells." in: The Journal of biological chemistry, Vol. 268, Issue 31, pp. 23072-80, 1993 (PubMed).

Faha, Harlow, Lees: "The adenovirus E1A-associated kinase consists of cyclin E-p33cdk2 and cyclin A-p33cdk2." in: Journal of virology, Vol. 67, Issue 5, pp. 2456-65, 1993 (PubMed).

Sherwood, Kung, Roitelman et al.: "In vivo inhibition of cyclin B degradation and induction of cell-cycle arrest in mammalian cells by the neutral cysteine protease inhibitor N-acetylleucylleucylnorleucinal." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, Issue 8, pp. 3353-7, 1993 (PubMed).

Darzynkiewicz, Gong, Juan et al.: "Cytometry of cyclin proteins." in: Cytometry, Vol. 25, Issue 1, pp. 1-13, 1997 (PubMed).

Gong, Traganos, Darzynkiewicz: "Expression of cyclins-B and cyclins-e in individual molt-4 cells and in stimulated human-lymphocytes during their progression through the cell-cycle." in: International journal of oncology, Vol. 3, Issue 6, pp. 1037-42, 2011 (PubMed).

Hosts (202), (62)
Reactivities (244), (95), (88), (27), (26), (16), (13), (13), (12), (2), (1), (1), (1)
Applications (177), (78), (71), (69), (53), (43), (40), (18), (14), (7), (3), (2), (1), (1)
Conjugates (7), (4), (4), (4), (4), (4), (4), (4), (4), (4), (4)
Epitopes (34), (27), (19), (13), (5), (4), (4), (4), (3), (3), (3), (3), (3), (3), (3), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Add to Basket

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Validation Images
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