- SourceMouse monoclonal antibody is produced from a hybridoma resulting from fusion of SP2/0 myeloma and B-lymphocytes obtained from a mouse immunized with Rituximab F(ab")2.
- IsotypeIgG1/kappa
- SpecificityRecognizes Rituximab specifically, no cross reactivity with other humanized antibodies.
- Purity
>95% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 years under sterile conditions after reconstitution.
Anti-Rituximab Antibodies (recommended for ADA assay) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development.Immobilized rituximab at 1 µg/mL, add increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 10% human serum) and then add biotinylated rituximab at 2 µg/mL. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 9.7 ng/mL.
Demonstration of the specificity of Anti-Rituximab Antibodies (Cat. No. RIB-Y36) to the rituximab.
Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development.Immobilized rituximab at 1 µg/mL, add increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 10% human serum) and then add biotinylated rituximab at 2 µg/mL. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 9.7 ng/mL.
Anti-Rituximab Antibodies (mouse IgG1, Cat. No. RIB-Y36) captured on CM5 chip via anti-mouse antibodies surface, can bind human rituximab with an affinity constant of 0.01 nM.
FACS analysis shows that the binding of rituximab to 293F overexpressing CD20 was inhibited by increasing concentration of Anti-Rituximab Antibodies (Cat. No. RIB-Y36). The concentration of rituximab used is 10 ng/ml. The IC50 is 0.025 μg/ml (Routinely tested).
- BackgroundRituxan is a genetically engineered chimeric murine/human monoclonal antibody directed against the CD20 antigen found on the surface of normal and malignant B lymphocytes. The antibody is an IgG1 kappa immunoglobulin containing murine light- and heavy-chain variable region sequences and human constant region sequences. Rituximab is composed of two heavy chains of 451 amino acids and two light chains of 213 amino acids.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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1. 血细胞与血清分离：
取人血液 1000ml,放置10min, 1000rpm离心20min .弃沉淀,留上清备用(沉淀为血细胞,上部为血清).
2. 乳糜粒分离：4000rpm 10°C离心10分钟,采用密度梯度离心
梯度液配置：离心管下部3/4容积加血浆,上部1/4容积加0.5MnaCl+0.3MEDTA,PH7.4 乳糜粒上浮,将乳糜粒吸出,留其余液体备用.
3. 血清蛋白分离：除去球蛋白,白蛋白及其它蛋白质.
①在用超滤除白蛋白，IgG中硫酸铵盐时，膜包该如何选择？选择几个？
②如用凝胶过滤来分级纯化血清中65%硫酸铵盐析出的白蛋白和纯化33%硫酸铵盐析出的IgG，其填充介质应选择什么？其洗脱缓冲液应用什么好？
下面有些关于这方面的数据，供参考：
①牛血清白蛋白：分子量：66210；分子形状：椭圆形；分子大小：
40Å*140Å；等电点：4.7；血浆中的含量：52.0g/L。
②IgG:分子量：15300；分子形状：球状；等电点：5.8—7.3;血浆中含量：2.0g/L。
③另外，我从书上看到说：凝胶过滤在分级方法中分辨率为中等，但对脱盐效果优良；流速较低，对分级每周期约≥8小时，对脱盐仅30分钟；适用于大规模纯化的最后步骤，在纯化过程的任何阶段均可进行脱盐处理，尤其适用于两种缓冲液交替时。
期待您的帮助，谢谢您。
1.以沉淀形式保存在高浓度的硫酸铵中。
2.添加50%甘油冷冻，尤其适用于酶类。
3.如果样品要用于生物学检测，避免使用防腐剂。在体内实验中不能添加防腐剂，而应当将样品分装成小份冷冻。
4.可使用无菌滤器以延长保存时间。
5.添加稳定剂，如甘油（5-20%）、血清白蛋白（10mg/ml）、配基（浓度取决于活性蛋白的浓度）以帮助维持生物活性。
6.避免反复冻融或冻干重溶解过程，这样很可能会降低生物活性。
7.一些冷沉淀蛋白会在4度沉淀出来，包括一些小鼠IgG3亚族的抗体，不能保存在4度冰箱中，可添加防腐剂保存在室温中。
