Anti- Rituximab (Anti-Idiotype) Antibody
Anti- Rituximab (Anti-Idiotype) Antibody Summary
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
![Detection of Rituximab antibody by Western Blot. Detection of Rituximab antibody by Western Blot.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_Western_Blot_21633.jpg)
Detection of Rituximab by Western Blot. Western blot shows Cetuximab, Rituximab, Brentuximab, and Gemtuzumab. PVDF membrane was probed with 0.5 µg/mL of Anti- Rituximab (Anti-Idiotype) Monoclonal Antibody (Catalog # MAB9630) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Rituximab at approximately 240 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.
![Anti-MS4A1/CD20 (Rituximab) binding to MS4A1/CD20-transfected HEK293 Human Cell Line is Blocked by Anti-Rituximab (Anti-Idiotype) Antibody. Anti-MS4A1/CD20 (Rituximab) binding to MS4A1/CD20-transfected HEK293 Human Cell Line is Blocked by Anti-Rituximab (Anti-Idiotype) Antibody.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_Block_Neutralize_21631.jpg)
Anti-MS4A1/CD20 (Rituximab) binding to MS4A1/CD20-transfected HEK293 Human Cell Line is Blocked by Anti-Rituximab (Anti-Idiotype) Antibody. In a functional flow cytometry test, Biotinylated Anti-MS4A1/CD20 (Rituximab) (Catalog # FAB9575B) binds to HEK293 human embryonic kidney cell line transfected with human MS4A1/CD20 (black dotted line). Binding is completely blocked (orange histogram) by 0.25 µg/mL of Anti-Rituximab (Anti-idiotype) Monoclonal Antibody (Catalog# MAB9630), but not by Normal Rabbit IgG (Catalog # MAB1050) (blue line). Cells were stained with APC-conjugated Streptavidin (Catalog # F0050).
![Detection of Anti-MS4A1/CD20 (Rituximab) on Human MS4A1/CD20 transfectants antibody by Flow Cytometry. Detection of Anti-MS4A1/CD20 (Rituximab) on Human MS4A1/CD20 transfectants antibody by Flow Cytometry.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_Flow_Cytometry_21630.jpg)
Detection of Anti-MS4A1/CD20 (Rituximab) on Human MS4A1/CD20 transfectants by flow cytometry. HEK293 human cell line transfected with (A) human MS4A1/CD20 or (B) irrelevant transfectants and eGFP were stained with Human anti-Human MS4A1/CD20 (Rituximab) Monoclonal Antibody (Catalog # MAB9575), then Anti-Rituximab (Anti-Idiotype) (Catalog # MAB9630) followed by APC-conjugated anti-rabbit IgG secondary antibody (Catalog # F0111).
![Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in Antibody in ELISA Capture Assay. Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in Antibody in ELISA Capture Assay.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_ELISA_21623.jpg)
Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in ELISA Capture Assay. Biotinylated biosimilars were serially diluted and incubated with Anti-Rituximab (Anti-Idiotype) Antibody (Catalog # MAB9630, green line) coated at 1 μg/mL on a 96-well polystyrene plate. No cross-reactivity with Brentuximab (black line) or biotinylated IgG control (orange line) was observed.
![Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in Direct Antibody in ELISA. Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in Direct Antibody in ELISA.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_ELISA_21646.jpg)
Specificity of Anti-Rituximab (Anti-Idiotype) Antibody in Direct ELISA. A panel of biosimilars was coated at 1 μg/mL in 96-well polystyrene plates. Anti-Rituximab (Anti-Idiotype) Antibody (Catalog # MAB9630, 333 ng/mL) detected Rituximab. 100% cross-reactivity with Tositumomab and Ibritumomab (95% or higher sequence identity with Rituximab) and no cross-reactivity with Cetuximab, Trastuzumab, Brentuximab, Gemtuzumab, Adalimumab, Solanezumab, Teprotumumab, and Aducanumab was observed.
![Anti-Rituximab Bridging Assay. Anti-Rituximab Bridging Assay.](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/antibody/Rituximab_MAB9630_ELISA_21627.jpg)
Anti-Rituximab Bridging Assay. The anti-idiotype antibody (catalog number MAB9630) directed against Rituximab was coated at 4 μg/mL in a 96-well polystyrene plate to capture the biosimilar Rituximab. Biotinylated anti-Rituximab (Anti-Idiotype) Antibody was used at 0.5 μg/mL as the detection antibody before incubation with Streptavidin-HRP. Other biosimilars did not bridge the anti-idiotype antibody, showing specificity of the anti-Rituximab (Anti-Idiotype) antibody.
![](https://aeroglobalimagesuat.blob.core.windows.net/images/datasheets/mab9630_anti-rituximab-anti-idiotype-mab-clone-2260a-other-162023102457.png)
NKs cells are potent killers in ADCC assays with Raji cells. 200x105 human NK cells were seeded from PBMCs and cultured for 13 days in plates/flasks using plate-bound Anti-Human NKp46 Monoclonal Antibody in ExCellerate Human NK Cell Expansion Media (Xeno-Free) plus cytokines (see expansion protocol, CCM032). Prior to setting up the ADCC assay, CD56+CD16+ levels were ascertained to be >30% on NKs using Human Fc gamma RIII (CD16) PerCP-conjugated Antibody (FAB2546C). Raji target cells were labeled with Mito Mark Green and Effectors/NKs were labeled with Janelia Fluor 646 (see FMC032 for details). Raji cells were preincubated with 10 ug/mL anti-CD20/Rituximab (Catalog # MAB9630) for 30 minutes at room temperature prior to setting up the ADCC assay with the NKs. 2:1 E:T ratios were used for the Raji/Rituximab model, and NKs and Raji cells were co-incubated for 2.5 hours at 37°C. The cells were then harvested, stained with antibodies for Human CD3 Alexa Fluor 405 and Human CD56 Alexa Fluor 700, and analyzed by flow cytometry. Importantly, non-NKs (JF 646-CD56- cells) are shown in the Mito Mark dye graph that depict killing. These results are representative of 3+ experiments.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Rituximab
The anti-Rituximab antibody is an anti-Idiotype antibody raised
against and selected for the antigenic determinants of the
variable region of Rituximab. The antibody has been tested against a
broad range of other biosimilars to ensure specificity. This highly
specialized anti-idiotypic antibody
can be used in a variety of pharmacokinetic (PK) assay formats. In addition to Rituximab, the anti-Idiotype antibody also detects derivatives of Rituximab (Ibritumumab and Tositumumab).
Product Datasheets
FAQs
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What is an anti-idiotype antibody?
An anti-idiotype antibody is specific for the unique variable (antigen binding) region of another antibody. We have anti-idiotype antibodies to commecially available therapeutic antibodies or biosimilars.
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