电话:
400-7060-959
传真:
+86 10 56315212-8813
电子邮件:
orders@antibodies-online.cn

HeLa Whole Cell Lysate (Nocodozole Stimulated)

WB Whole Cell Lysate Cell Lysate HeLa Cells Nocodozole Stimulated 适用: 人
产品编号 ABIN964038
发货至: 中国
  • 宿主
    资源
    人细胞
    应用范围
    Western Blotting (WB)
    特异性
    The cells were grown in DMEM supplemented with 10% FBS (Fetal Bovine Serum). Cells were treated with 3 µg/ml nocodazole for 1 h. Cells were washed in PBS and incubated on ice in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid and 0.1% SDS to lyse the cells. Protein integrity is ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 µM Aprotinin, 5 µM Bestatin, 1.5 µM E-64, 2 µM Leupeptin Hemisulfate and 1 µM Pepstatin A). Phosphatase inhibitors sodium fluoride, sodium orthovanadate, sodium pyrophosphate and -glycerophosphate were also added. Cell debris was removed by centrifugation. Protein concentration was determined by Lowry assay using a commercially available kit. The protein concentration was adjusted to 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added.
    产品特性
    Cell Line: HeLa - Human epidermoid carcinoma
    Induction: Nocodazole (3 µg/ml)
    过滤
    Sterile filtered
    Lysed Cells
    HeLa Cells
    Lysate Type
    Cell Lysate
    Lysate Fraction
    Whole Cell Lysate
    Lysate Treatment
    Nocodozole Stimulated
  • 应用备注
    Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added). Heat lysate to 95° C for 5 minutes and rapidly cool.  If dissociating conditions are desired, add reducing agent prior to heating.  The recommended loading volume per lane is 10-20 µl depending on the size format of your gel.
    说明

    Lysate Fractionation: Whole Cell Lysate
    Lysate Stimulation: Nocodozole
    Lysate Tissue Culture: Tissue Culture

    限制
    仅限研究用
  • 状态
    Liquid
    浓度
    1.0 mg/mL
    缓冲液
    1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
    注意事项
    Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added). Heat lysate to 95 °C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior to heating. The recommended loading volume per lane is 10-20 µl depending on the size format of your gel.
    储存条件
    -80 °C
    有效期
    3 months
  • 背景
    Ready-to-use whole cell lysates produced are derived from cell lines or tissues using highly refined extraction protocols to ensure exceptionally high quality, protein integrity and lot-to-lot reproducibility.  All extracts are tested by SDS-PAGE using 4-20% gradient gels and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.
You are here:
客服