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pH-sensitive lgG labeling reagents (Deep Red) STAR

价格 4,400.00 供应商现货 : 3-5个工作日
货号 UA079026
规格
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产品规格
  • 分子量

    31kDa

  • 缓冲体系

    PBS, PH7.4 with 5% trehalose is added as protectants

  • 溶解方法


    1. Before opening the tube cap, centrifuge the sample tube at 5000g for 3-5min at room temperature to ensure the lyophilized sample to settie down at the bottom of the tube.


    2. Dissolved lyophilized protein sample in sterile water based on the recommended volume 10uL(10ug package).


    3. After adding sterile water, cover the lid and mix them bygently tapping the tube for 5-10 times. Note: Do not vortex or vigorously pipette sample.

  • 储存条件

    · 12 months from date of receipt, lyophilized powder stored at -20 to -80℃.
    · 1 week, 2 to 8℃ under sterile conditions after reconstitution.

背景介绍
  • Antibody internalization labeling probe. It can bind to candidated antibodies to produce soluble complex that only fluoresce in acidic environments for the study of antibody internalization.
    The pH-sensitive Labeling probe binds to your primary antibodies via Fc binding protein to a pH-dependent fluorescent molecular. This fluorescent complex reporter will increase intensity as the pH of its surroundings becomes more acidic, as evident when exposed to the environment inside a cell. The internalized antibodies are detected by measuring fluorescence intensity of the cells.

    This product can becan be used for human IgG1, IgG2, IgG3, IgG4, rabbit IgG, mouse IgG1, IgG2a, IgG2b and IgG3 and can be detected with Flow cytometry Cy5 filter.

操作步骤
  • 1.UA079026 与抗体孵育
    1.1 使用去离子水,将冻干粉形式的UA079026根据溶解方法进行复溶。
    1.2 用细胞培养基制备足够体积的 4X待测抗体工作溶液(受试浓度的四倍,受试浓度根据前期流式条件摸索)。例如:过表达细胞2μg/ml可能是抗体的良好起始测试浓度,因此 4X工作溶液为 8μg/ml;肿瘤细胞建议参考已验证浓度,或者是200nM-300nM梯度稀释摸索最佳用量。
    1.3 用细胞培养基制备足够体积的4X UA079026工作溶液。待测抗体与UA079026 摩尔比 1:2。
    1.4 将4X待测抗体工作溶液和4X UA079026工作溶液按 1:1 的体积比混合。室温避光孵育15min- 1h,得到 Ab- UA079026 复合物2X工作液。
    2. Ab-UA079026复合物与细胞孵育
    2.1细胞处理:收集、洗涤细胞,使用完全培养基调整细胞浓度,悬浮细胞 1-2×105 cells/mL 的浓度,贴壁细胞 0.5-1×105cells/mL 的浓度,96 孔板每孔加入 100μL 细胞悬液。
    2.2 孵育:每孔加入 100μL Ab- UA079026 复合物 2X 工作液,37℃ 5% CO2 培养箱中培养。
    3. 流式检测
    培养 18-24h 后 (可根据抗体实际内吞时间调整) ,收集细胞进行流式检测抗体的内吞效果,使用APC或Cy5通道检测,激发波长(Ex):640nm,发射波长(Em):655nm。

  • 生物活性

    • Fig1. The fluorescent intensity from antibody-probe protein complex internalization into cell is specific for B7-H3 on Hela cell.

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