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Biotinylated Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Y45)

表达区间及表达系统(Source)

Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 is produced from a hybridoma resulting from fusion of SP2/0 myeloma and B-lymphocytes obtained from a mouse immunized with FMC63 scFv. It is the biotinylated form of Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (FM3-Y45).

亚型(Isotype)

IgG1/kappa

特异性(Specificity)

Specifically recognizes the antigen-recognition domain of FMC63 derived CARs.

标记(Biotinylation)

The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A specially optimized long-spacer biotin reagent (13.5 angstroms) is used in this product to minimize potential steric hindrance.

蛋白标记度(Biotin:Protein Ratio)

The biotin to protein ratio is 0.5-2 as determined by the HABA assay.

应用说明(Application)

Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression).

制剂(Formulation)

Lyophilized from 0.22 μm filtered solution in PBS, 0.5% BSA, pH7.4. Normally trehalose is added as protectant before lyophilization.

重构方法(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.

存储(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:

  1. -20°C to -70°C for 12 months in lyophilized state;
  2. -70°C for 6 months under sterile conditions after reconstitution.

活性(Bioactivity)-FACS

Biotinylated Human FACS

2e5 of Anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) Biotinylated Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-BY45) and isotype control respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).

Protocol

 

背景(Background)

FMC63 is an IgG2a mouse monoclonal antibody specific for CD19, which is a target for the immunotherapy of B lineage leukaemias and lymphomas. FMC63 scFv is the most commonly used ectodomain component of CD19-specific CARs. So far, most of reported CART19 trials contain the anti-CD19 scFv derived from FMC63, including the two FDA-approved CARs Kymriah and Yescarta.

 

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