Cyclin B1 抗体 (CCNB1)

Details for Product anti-CCNB1 Antibody No. ABIN967429, 供应商: Log in to see
抗原
  • ccnb
  • cycb
  • cb267
  • cycb1
  • wu:fa19g04
  • wu:fb16d01
  • wu:fb16e07
  • wu:fi21c01
  • ccnb1
  • MGC53596
  • CCNB
  • Ccnb1-rs1
  • Ccnb1-rs13
  • CycB1
  • Cycb-4
  • Cycb-5
  • Cycb1-rs1
  • cyclin B1
  • ccnb1
  • ccnb1.2
  • CCNB1
  • Ccnb1
  • LOC100338989
适用
人, 小鼠
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小鼠
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克隆类型 (克隆位点)
单克隆 ()
标记
This Cyclin B1 antibody is un-conjugated
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应用范围
BioImaging (BI), Flow Cytometry (FACS), Immunoprecipitation (IP), Western Blotting (WB)
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选项
Supplier
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Supplier Product No.
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品牌 BD Pharmingen™
免疫原 Recombinant Human Cyclin B1
克隆位点 GNS-11
亚型 IgG2a
交叉反应 小鼠
产品特性 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.
纯化方法 The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
别名 Cyclin B1 (CCNB1 Antibody 摘要)
背景 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.
分子量 62 kDa
研究领域 Cancer, Cell Cycle
途径 Cell Division Cycle, AMPK Signaling, Mitotic G1-G1/S Phases, M Phase
应用备注 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.
说明

Related Products: ABIN967389, ABIN968537

限制 仅限研究用
状态 Liquid
浓度 0.5 mg/mL
缓冲液 Aqueous buffered solution containing ≤0.09 % sodium azide.
储存液 Sodium azide
注意事项 This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
储存条件 4 °C
储存方法 Store undiluted at 4°C.
厂商提供的图像
Western Blotting (WB) image for anti-Cyclin B1 (CCNB1) antibody (ABIN967429) Western blot analysis of Cyclin B1. Lysate from Jurkat T cells was probed with anti- ...
Immunofluorescence (IF) image for anti-Cyclin B1 (CCNB1) antibody (ABIN967429) Immunofluorescent staining of HeLa (ATCC CCL-2) cells. Cells were seeded in a 96 well...
背景 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).

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).

Keyomarsi, Pardee: "Redundant cyclin overexpression and gene amplification in breast cancer cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, Issue 3, pp. 1112-6, 1993 (PubMed).

Minow: "The role of families in medical decisions." in: Utah law review, Vol. 1991, Issue 1, pp. 1-24, 1993 (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).

Hoffmann, Clarke, Marcote, Karsenti, Draetta: "Phosphorylation and activation of human cdc25-C by cdc2--cyclin B and its involvement in the self-amplification of MPF at mitosis." in: The EMBO journal, Vol. 12, Issue 1, pp. 53-63, 1993 (PubMed).

Jessus, Beach: "Oscillation of MPF is accompanied by periodic association between cdc25 and cdc2-cyclin B." in: Cell, Vol. 68, Issue 2, pp. 323-32, 1992 (PubMed).

Pines, Hunter: "Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport." in: The Journal of cell biology, Vol. 115, Issue 1, pp. 1-17, 1991 (PubMed).

Dunphy, Brizuela, Beach, Newport: "The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis." in: Cell, Vol. 54, Issue 3, pp. 423-31, 1988 (PubMed).

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