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Product Details
Source
Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag™ (Y45) is a neutralizing mouse monoclonal antibody recombinantly expressed from human 293 cells (HEK293). Biotinylation of this antibody is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Application
Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression).
Clone
Y45
Species
Mouse
Isotype
Mouse IgG1 & | kappa
Specificity
Specifically recognizes the antigen-recognition domain of FMC63 derived CARs.
Immunogen
Recombinant FMC63 scFv derived from HEK293 cells
Conjugate
Biotin
Isotype Control
The isotype control is sold separately; please refer to Cat. No. DNP-BM190 for product information.
Recommended Dilution
1:50
Formulation
Lyophilized from a 0.22 μm-filtered solution in PBS (pH 7.4) containing 0.5% BSA, with trehalose as protectant.
Please contact us for customized product forms or formulations.
Reconstitution
Please refer to the Certificate of Analysis (CoA) for specific instructions.
For best performance, we strongly recommend following the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored in a lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 24 months in lyophilized state;
- -70°C for 12 months under sterile conditions after reconstitution.
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Performance Data
Bioactivity-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 Antibody, Mouse IgG1, Avitag (Cat. No. FM3-BY54) and isotype control respectively, washed and then followed by PE-SA and analyzed with FACS (QC tested).
Protocol
Non-specificity of Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag (Cat. No. FM3-BY54) binding to CD3+ cells present in human PBMC. Human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag (2 μL of the antibody stock solution corresponds to labeling of 5e5 cells in a final volume of 100 µL), washed and then analyzed with FACS. Both FITC and PE positive signals was used to evaluate the non-specific binding activity to human CD3+ cells (QC tested).
Protocol
FACS Analysis of Blood Samples
After Fc receptor blockade, whole blood samples (Figure A) and whole blood spiked with CAR-T cells (Figure B) were first stained with Biotinylated Monoclonal Anti-FMC63 Antibody, Mouse IgG1, Avitag™ (Cat. No. FM3-BY54). Following incubation with Streptavidin Protein-PE (Cat. No. STN-NP117) and 7-AAD staining, the cells were analyzed using FCS Express 7 software (Routinely tested). CAR-T cells were identified as PE-positive events within the live CD45+CD3+ lymphocyte population.
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