为了使您在Abcam官网的浏览体验更顺畅,请使用最新版本的浏览器比如 Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
致电 +86 21 2070 0500 或 联系我们

  • 我的账户
  • 退出
登录 或 注册

欢迎光临

登录 或

您还没有帐户?

注册
我的购物篮
快速订购
Abcam homepage

  • 研究用途的产品
    按产品类型
    一抗
    二抗
    免疫测定试剂盒与试剂
    细胞与组织成像工具
    细胞与生化测定
    蛋白质与肽
    按产品类型
    蛋白质组学工具
    激动剂、活化剂、拮抗剂与抑制剂
    细胞株和裂解液
    多重miRNA检测
    多因子分析试剂盒
    按研究领域
    癌症
    心血管
    细胞生物学
    表观遗传学
    代谢
    发育生物学
    按研究领域
    免疫学
    微生物学
    神经科学
    信号转导
    干细胞
  • 定制化产品 & 合作关系
    定制化产品 & 合作关系

    定制化产品和商务合作加速您的诊断和治疗项目

    定制化产品

    与我们成为伙伴

  • 技术支持
    支持中心

    获取研究建议和支持

    查看支持中心

    Protocols

    一步一步完成您的实验

    查看Protocols

  • 活动通知
    • 会议年历
    • 肿瘤研究
    • 心血管研究
    • 染色质及细胞核信号转导研究
    • 免疫学
    • 神经生物学
    • 干细胞研究
    • 商品展览
    • 实验技术视频
    查看最新活动

    活动说明、摘要提交、演讲者、注册方式及更多信息

    查看全球活动安排

  • 通路
    细胞信号转导通路

    查看所有通路

    查看交叉通路

Induction of apoptosis in cells

Related

  • All apoptosis resources
    • Apoptosis analysis ebook
      • Apoptosis pathway card
        • Apoptosis assay kits guide
          • Role of caspases in apoptosis
            • Mitochondrial function in cell death | Webinar

              Procedure for biological and chemical induction of apoptosis in cells. 

              Print this protocol.

              ​​​​​​​​​​​​​​​​​​​​​​Apoptosis may be induced in experimental systems through a variety of methods. In general, methods to induce apoptosis can be divided into two categories: biological induction and chemical induction.

              ​​Biological induction of apoptosis

              ​​​Activation of Fas or TNF receptors by their respective ligands, or by cross-linking with an agonist antibody, induces apoptosis of Fas- or TNF receptor-bearing cells. Here we describe a general protocol to induce apoptosis using an anti-Fas receptor (anti-CD95) monoclonal antibody (mAb) in Jurkat cells. The mouse monoclonal anti-Fas antibody [UT-1] (ab11881) can be used for apoptosis indication.

              1. Grow Jurkat cells in RPMI-1640 containing 10% fetal bovine serum (FBS) in a humidified 5% CO2 incubator at 37°C.
              2. Harvest exponentially growing cells at a concentration of 105 cells/mL by centrifugation at 300–350 x g for 5 min. Resuspend cells in fresh medium to a final concentration of 5 x 105 cells/mL.
              3. Add anti-Fas (anti-CD95) mAb to the appropriate concentration(for ab11881, use a final concentration of 2–20 mg/mL. Incubate for 2–4 hr in a 37°C incubator. For a negative control, incubate untreated cells (without anti-Fas/CD95 antibody) under the same conditions.
              4. Harvest the cells by centrifugation at 300–350 x g for 5 min.
              5. Remove all medium and resuspend cells in PBS.
              6. Repeat centrifugation step and resuspend cells in PBS to 1.5 x 106 cells/mL.
              7. Proceed to detect apoptosis using your method of choice.

              Chemical induction of apoptosis

              Apoptosis inducers act on several apoptosis-related proteins to promote apoptotic cell death. Depending on the agent selected and the concentrations used, apoptotic events can be detected between 8–72 h post-treatment. However, not all reagents will affect a particular cell line in the same way. 

              These are general guidelines for inducing cellular damage with chemical agents that will lead to apoptosis.

              1. Set up your cells for treatment: inoculate adherent cells into 10 cm2 tissue culture dishes or suspension cells into T75 flasks at a concentration of ~106 cells/mL. Prepare as many samples as necessary to ensure you can detect the induction of apoptosis at the relevant time. One dish/flask will be used as negative control for non-induced cells.
              2. Add cellular damaging agents at the recommended concentrations to induce apoptosis.

                For instance, the list below suggests final concentrations of cellular damaging agents that can be used for induction of apoptosis through p53-dependent G1 arrest:

                AgentConcentration
                Doxorubicin hydrochloride (ab120629)0.2 µg/mL (25 µg/mL stock
                prepared in H2O)
                Staurosporine (ab120056)1 µM (1 mM stock prepared in
                DMSO)
                Etoposide (ab120227)1–10 µM (1 mM stock prepared
                in DMSO)
                Camptothecin (ab120115)2–10 µM (1 mM stock prepared
                in DMSO)
                Paclitaxel (ab120143)50–100 nM (stock prepared in
                DMSO)
                Vinblastine (ab145649)60 nM (stock prepared in
                ​methanol)
                ​Add appropriate volume of buffer or solvent to the negative control.

                ​As a further control, inhibitors to the different pathways can be included:

                AgentConcentration
                Z-VAD-FMK (ab120382)50 µM (stock prepared in DMSO)
              3. Harvest cells at different times (i.e. 8, 12, 16, 24, 48, 72 h) after the addition of the cellular damaging agent.
              4. For apoptosis detection: Collect cells and detect apoptosis using your method of choice.
              5. For examination of apoptotic proteins: Collect all cells. Prepare cell lysates for either western blot detection or immunoprecipitation using your method of choice. Always compare levels of apoptotic proteins with control treated cells to confirm induction.
              View our apoptosis detection assays
              通过邮件收到学术资源和促销信息 注册
              研究类别(A-Z)
              • 肿瘤研究
              • 心血管研究
              • 细胞生物学
              • 发育生物学
              • 染色质及细胞核信号转导研究
              • 免疫学
              • 代谢
              • 微生物学
              • 神经生物学
              • 信号转导研究
              • 干细胞研究
              产品种类(A-Z)
              • 一抗
              • 二抗
              • 生化试剂
              • 亚类对照
              • 流式细胞仪多色选择器
              • 试剂盒
              • 内参对照
              • 细胞裂解液
              • 多肽
              • 蛋白酶
              • 组织切片
              • 蛋白标签及细胞标记物
              • 相关试剂及工具
              帮助与支持
              • 技术支持
              • 技术FAQ
              • 实验步骤与建议
              • 购买FAQ
              • RabMAb 产品
              • 培训课程
              • 按靶标浏览
              企业
              • 企业资讯
              • 投资者关系
              • 公司新闻
              • 招贤纳士
              • 关于我们
              活动通知
              • 商品展览
              • 会议
              全球网站
              • abcam.com
              • abcam.co.jp
              QR code for Weechat

              微信扫码 关注我们

              沪ICP备14046373号-1

              沪公网安备 31011502007413号

              • 销售条款
              • 网站使用条款
              • Cookie政策
              • 隐私权政策
              • 法律
              © 1998-2023 艾博抗(上海)贸易有限公司版权所有