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Product Details
Source
APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) is a neutralizing antibody produced via site-specific conjugation of APC to Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 under optimal conditions with a proprietary technology.
Application
Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression). Please note that this product is NOT compatible to streptavidin detection system.
Clone
Y45
Species
Mouse
Isotype
Mouse IgG1 | Mouse Kappa
Specificity
Specifically recognizes the antigen-recognition domain of FMC63 derived CARs.
Immunogen
Recombinant FMC63 scFv derived from HEK293 cells
Conjugate
APC
Excitation Wavelength: 640 nm
Emission Wavelength: 661 nm
Recommended Dilution
1:50
Formulation
Lyophilized from a 0.22 μm-filtered solution in PBS (pH 7.4) containing 0.03% ProClin 300 and 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 protect from light and 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 after reconstitution;
- 2-8°C for 12 months after reconstitution.
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Performance Data
Bioactivity-FACS
5e5 of anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) of APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Cat. No. FM3-AY54A1) and isotype control antibody respectively. APC signal was used to evaluate the binding activity (QC tested).
Protocol
Non-specificity of APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Cat. No. FM3-AY54A1) binding to CD3+ cells present in human PBMC. Human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (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 APC positive signals was used to evaluate the non-specific binding activity to human CD3+ cells (QC tested).
Protocol
Binding activity of the APC-Labeled Monoclonal Anti-FMC63 Antibody before and after adding 0.03% Proclin was evaluated in the above FACS analysis. The result shows that APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Site-specific conjugation) (Cat. No. FM3-AY54P1) and APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Site-specific conjugation) (0.03% Proclin) (Cat. No. FM3-AY54A1) have almost the same binding activity against anti-CD19 CAR-293 cells.
Protocol
Evaluation of CAR expression
FACS Analysis of Non-specific binding to Human whole blood
Non-specificity of APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Site-specific conjugation) (0.03% Proclin)(Cat. No. FM3-AY54A1) binding to CD3+ cells present in human whole blood. 100 μl of human whole blood were simultaneously stained with APC/Cyanine7 anti-human CD3 Antibody and APC-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (2 μL of the antibody stock solution in a final volume of 100 µL), compared with isotype control antibody and positive control antibody. Both APC/Cyanine7 and APC positive signals was used to evaluate the non-specific binding activity to human CD3+ cells.
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