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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.
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 FITC or AF488 filter.
1. UA070122 与抗体孵育
1.1 使用去离子水，将冻干粉形式的 UA070122 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：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。
1.3 用细胞培养基制备足够体积的4X UA070122 工作溶液。待测抗体与 UA070122 摩尔比 1:2。
1.4 将4X待测抗体工作溶液和4X UA070122 工作溶液按 1:1 的体积比混合。室温避光孵育 1h，得到 Ab- UA070122 复合物2X工作液。
2. Ab- UA070122 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 0.5-1×10^5cells/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- UA070122 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，收集细胞进行流式检测抗体的内吞效果，使用 FITC 或AF488 通道检测
Avoid vigorous vortex after reconstitution.
Avoid repeated freeze-thaw cycles.
生物活性JSON
Fig1. The fluorescent intensity from antibody-probe protein complex internalization into cell is specific for B7-H3 on Hela cell.
Fig2. Comparison of antibody for B7-H3 internalization effects between UA070122 and UA070080 on Hela cells.
长期稳定性和孵育时间的影响
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