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細胞增殖和毒性檢測-BrdU法

更新時間:2012-12-28 09:23:53點擊次數:4196次字號:T|T
◆ BrdU ◆ Mouse BrdU Monoclonal Antibody ◆ Anti-mouse Secondary Antibody PART 1:5-溴脫氧尿嘧啶核苷(BrdU) 產品信息: 產地 產品名稱 貨號 規格 價格(元) 促銷價(元) Sigma BrdU B5002 100 mg 454.00 328.00 Sigma BrdU B5002 1 g 1224.00 958.00 化學特性: 名稱:(5-溴脫氧尿嘧啶核苷)BrdU 結構 別名:


     BrdU

     Mouse BrdU Monoclonal Antibody

     Anti-mouse Secondary Antibody

    PART 15-溴脫氧尿嘧啶核苷(BrdU

    產品信息:


    產地

    產品名稱

    貨號

    規格

    價格(元)

    促銷價(元)

    Sigma

    BrdU

    B5002

    100 mg

    454.00

    328.00

    Sigma

    BrdU

    B5002

    g

    1224.00

    958.00


    化學特性:


    名稱:5-溴脫氧尿嘧啶核苷)BrdU

    結構

    別名:Br-dU, BUdR, 5-BrdU, 5-Bromodeoxyuridine,5-Bromo-2-Deoxyuridine

    CAS. No.59-14-3

    分子式:C9H11BrN2O5

    分子量:307.1

    外觀:粉末

    保存方法:-20

    溶解:Sigma QC 測試Brdu1 M氨溶液中的溶解度為50 mg/ml。Brdu也可溶于水,溶解性高達10 mg/ml。也可溶于DMF, DMSO和水(加熱)(50-100 mg/ml)。


    產品描述:

    5-溴脫氧尿嘧啶核苷(BrdU)是胸腺嘧啶的衍生物,常用于標記活細胞中新合成的DNA,可代替胸腺嘧啶選擇性整合到復制細胞中新合成的DNA中(細胞周期S期)。這種摻入可以穩定存在,隨著DNA復制進入子細胞中。BrdU 特異性抗體可以用于檢測BrdU的摻入,從而判斷細胞的增殖能力。Brdu與抗體的結合需要DNA變性,通常用酸或加熱的方法進行處理。Brdu在標記濃度范圍內既不具有放射活性也沒有骨髓毒性,因此也常用于研究腫瘤細胞的體內增殖狀況。

    產品應用:

    v         體外實驗:

    1)   為檢測EBs增殖,在培養基中加入30 μM BrdU[1]。

    2)   為檢測ShhEGL細胞增殖的影響,用重組的Shh-N培養細胞,其中含有10 μM BrdU[2]。

    3)   為檢測小鼠胚胎成纖維細胞(MEFs)的增殖,細胞在含有10 μM BrdU的培養基中孵育4h,anti-BrdU 單克隆抗體(1:10)孵育1h, wesern blottin檢測[3]。

    4)   為檢測大鼠SMCs細胞增殖,用含有0.5% FCSDMEM培養基培養細胞,加入10 μM BrdU [4]。

    v         體內實驗:

    1)   在小鼠全組織包埋實驗中,給小鼠腹腔注射100 μg/gm體重的BrdU,用1:200 anti-BrdU抗體孵育48-72h[5]。

    2)   為標記有絲分裂活躍的細胞,給大鼠腹腔注射50 mg/kg BrdUPBS溶解)。用抗BrdU小鼠單克隆抗體(1:100)孵育組織切片[6]。

    3)   為檢測大鼠腦組織中BrdU標記的核酸,給hx 大鼠每天腹腔注射50 mg/kg 體重的BrdU,用小鼠anti-BrdU抗體(1:400)孵育樣品[7]。

    參考文獻:

    [1] Carmeliet, P., et al. Role of HIF-1α in hypoxiamediated apoptosis, cell proliferation and tumour angiogenesis. Nature, 394: 485-490(1998).

    [2] Wallace, V.A., et al. Purkinje-cell-derived Sonic hedgehog regulates granule neuron precursor cell proliferation in the developing mouse cerebellum. Current Biology, 9:445–448(1999).

    [3] Jacobs, J.L., et al. The oncogene and Polycombgroup gene bmi-1regulates cell proliferation and senescence through the ink4a locus. Nature, 397(14): 164-168(1999).

    [4] Janssens, S., et al. Human Endothelial Nitric Oxide Synthase Gene Transfer Inhibits Vascular Smooth Muscle Cell Proliferation and Neointima Formation After Balloon Injury in Rats. Circulation. 97:1274-1281(1998).

    [5] Doetsch, F., et al. Lack of the Cell-Cycle Inhibitor p27Kip1 Results in Selective Increase of Transit-

    Amplifying Cells for Adult Neurogenesis. The Journal of Neuroscience, 22(6): 2255-2264 (2002).

    [6] Kuhn, H.G., et al. Epidermal Growth Factor and Fibroblast Growth Factor-2 Have Different Effects on Neural Progenitors in the Adult Rat Brain. The Journal of Neuroscience, 17(15): 5820- 5829(1997).

    [7] Maria, A.I., et al. Peripheral Infusion of IGF-I Selectively Induces Neurogenesis in the Adult Rat Hippocampus. The Journal of Neuroscience, 20(8):2896-2903(2000).

    PART 2Mouse BrdU Monoclonal Antibody

    產品信息(現貨供應):


    產地

    產品名稱

    規格

    促銷價(元)

    DSHB

    G3G4(mouse anti-BrdU)

    200 ul

    1200.00


    抗體信息:


    Immunogen

    BrdU

    Isotype

    Mouse IgG1, kappa light chain  

    Reactivity

    All species

    Physical State

    Liquid

    Application

    IHC:2-5μg/ml;WB:0.2-0.5μg/ml;

    IF,IP(Use at an assay dependent dilution)

    Storage

    4 for short term storage (2 weeks).

    Recommend to aliquot and store at -20 for long term storage. Prevent from freeze-and-thaw cycles.


    引用文獻:

    [1] George-Weinstein, M., Foster, R.F., Gerhardt, J.V., and Kaufman, S.J. (1993). In vitro and in vivo expression of 7 integrin and desmin define the primary and secondary myogenic lineages. Dev. Biol. 156, 209-229.

    [2] Dominov, J.A., Dunn, J.J., and Miller, J.B. (1998). Bcl-2 expression identifies an early stage of myogenesis and promotesclonal expansion of muscle cells. J. Cell Biol. 142, 537-544.

    [3] Orris, A.S., Banicky, L.C., and Wiens, D.J. (1999). Isotretinoin alters morphology, polarity, and motility of neural crestcells in culture. Reprod. Tox. 13(1), 45-52.

    [4] Vicaro-Abejon, C., Yusta-Boyo, M.J., Fernandez-Moreno, C., and de Pablo, F. (2003). Locally born olfactory bulb stem cells proliferate in response to insulin-related factors and require endogenous insulin-like growth factor-I for differentiation into neurons and glia. J. Neurosci. 23(3), 895-906.

    [5]Yusta-Boyo, M.J., Gonzalez, M.A., Pavon, N., Martin, A.B., de la Fuente, R., Garcia-Castro, J., de Pablo, F., Moratalla, R.,

    [6]Bernad, A., and Vicario-Abejon, C. (2004). Absence of hematopoiesis from transplanted olfactory bulb neural stem cells.Eur. J. Neurosci. 19, 505-512.

    [7]Garelli, A., Rotstein, N.P., and Politi, L.E. (2006). Docosahexaenoic acid promotes photoreceptor differentiation without altering Crx expression. Invest. Ophthalmol. Vis. Sci. 47(7), 3017-3027.

    [8]Otaegi, G., Yusta-Boyo, M.J., Vergano-Vera, E., Mendez-Gomez, H.R., Carrera, A.C., Abad, J.L., Gonzalez, M., de la Rosa, E.J.,

    [9]Vicaro-Abejon, C., and de Pablo, F. (2006). Modulation of the PI 3-kinase-Akt signalling pathway by IGF-I and PTEN regulates the differentiation of neural stem/precursor cells. J. Cell Sci. 119, 2739-2748.

    [10]Vergano-Vera, E., Yusta-Boyo, M.J., de Castro, F., Bernad, A., de Pablo, F., and Vicario-Abejon, C. (2006). Generation of GABAergic and dopaminergic interneurons from endogenous embryonic olfactory bulb precursor cells. Development 133,4367-4379.

    [11]Francisco-Morcillo, J., Hidalgo-Sánchez, M., Martín-Partido, G., (2006). Spatial and temporal patterns of proliferation and differentiation in the developing turtle eye. Brain Res. 1103, 32-48.

    [12]Shibasaki, K., Takebayashi, H., Ikenaka, K., Feng, L., and Gan, L. (2007). Expression of the basic helix-loop-factor Olig2 in the developing retina: Olig2 as a new marker for retinal progenitors and late-born cells. Gene Expr. Patterns 7, 57-65.

    [13]Tamiya, S., Liu, LH, and Kaplan, H.J. (2010). Epithelial-Mesenchymal transtition and proliferation of retinal pigment epithelial cells initiated upon loss of cell-cell contact. Invest. Ophthalmol. Vis. Sci. 51(5), 2755-2763.

    [14] Gerbe, F., van Es, J.H., Makrini, L., Brulin, B., Mellitzer, G., Robine, S., Romagnolo, B., Shroyer, N.F., Bourgaux, J.-F., Pignodel, C., Clevers, H., and Jay, P. (2011). Distinct ATOH1 and Neurog3 requirements define tuft cells as a new secretory cell type in the intestinal epithelium. J. Cell Biol. 192(5), 767-780.

    [15] Martinez-de Luna, R.I., Kelly, L.E., and El-Hodiri, H.M. (2011). The retinal homeobox (Rx) gene is necessary for retinal regeneration. Dev. Biol. 353, 10-18.

    Part 3Anti-mouse secondary antibody

    Jackson ImmunoResearch Inc.是美國著名的二抗生產商。JIR公司的二抗以種類豐富(物種齊全,標記多樣,應用廣泛),性價比高,供貨迅捷而深受國際科研學者和工業客戶的青睞。JIR公司不僅堅持提供傳統標記的二抗(如酶標記,FITC,羅丹明,Cy系列等),而且始終與最前沿的染料生產商合作,以最快的速度供應新型染料標記二抗。目前市場上最受歡迎,口碑最好的莫屬DyLightTMAlexa Fluor?系列熒光染料,從客戶需求和公司發展戰略出發,JIR公司將在繼續提供部分DyLightTM標記二抗的基礎上,著重推出Alexa Fluor?熒光標記二抗。作為DyLight熒光染料的完美替代物,Alexa Fluor?同樣具有以下優點甚至表現更好:熒光強度高,光耐受性(抗淬滅性)強,水溶性好,儲存不會產生沉淀,pH值不敏感,儀器兼容性廣。另外從藍光到遠紅外范圍,能覆蓋多種顏色且分辨率高,非常適合多標實驗。

    山羊抗小鼠酶標二抗:


    貨號

    名稱

    原裝價格

    分裝價格

    115-035-003

    Peroxidase–conjugated Goat Anti-Mouse IgG (H+L) 山羊抗小鼠

    1392/2.0 ml

    200/100μl

    115-055-003

    AP–conjugated Goat Anti-Mouse IgG (H+L) 山羊抗小鼠

    1824/1.0 ml

    280/100μl


    山羊抗小鼠熒光二抗:


    貨號

    名稱

    原裝價格

    分裝價格

    115-165-003

    Cy? 3-conjugated Goat Anti-Mouse IgG(H+L) 山羊抗小鼠

    1392/2.0 mg

    200/100μl

    115-095-003

    FITC–conjugated Goat Anti-Mouse IgG(H+L) 山羊抗小鼠

    1088/2.0 mg

    180/100μl

    115-025-003

    TRITC)–conjugated Goat Anti-Mouse IgG(H+L) 山羊抗小鼠

    1088/2.0 mg

    180/100μl

    115-295-003

    Rhodamine Red?-X-conjugated Goat Anti-Mouse IgG(H+L)

    1088/2.0 mg

    180/100μl

    115-545-003

    Aleax Flour 488- conjugated Goat Anti-Mouse IgG(H+L)

    1456/1.5 mg

    220/100μl

    115-585-003

    Aleax Flour 594- conjugated Goat Anti-Mouse IgG(H+L)

    1456/1.5 mg

    220/100μl

    115-605-003

    Aleax Flour 647- conjugated Goat Anti-Mouse IgG(H+L)

    1456/1.5 mg

    220/100μl

    115-475-003

    DyLight 405- conjugated Goat Anti-Mouse IgG(H+L)

    1600/1.5 mg

    220/100μl


    遠紅外-紅外山羊抗小鼠熒光二抗(熒光WB,與LiCor Odyssey imaging systems兼容):


    貨號

    名稱

    原裝價格

    分裝價格

    115-625-146

    Alexa Fluor?680 goat anti-mouse(min X Hu, Bov, Hrs, Rb, Sw Sr Prot)山羊抗小鼠

    2064/0.5mg

    320/100ul

    115-655-146

    Alexa Fluor?790 goat anti-mouse(min X Hu, Bov, Hrs, Rb, Sw Sr Prot)山羊抗小鼠

    2064/0.5mg

    320/100ul


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