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Product Details
Source
Alexa Fluor 647-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) is a neutralizing antibody produced via conjugation of AF647 to Monoclonal Anti-FMC63 Antibody, Mouse IgG1 under optimal conditions with a new generation site-specific technology under Star Staining labeling platform.
Application
Flow Cytometry (Evaluation of Anti-CD19 (FMC63 scFv) CAR Expression).
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
AF647
Excitation Wavelength: 640 nm
Emission Wavelength: 672 nm
Recommended Dilution
1:50
Formulation
Lyophilized from a 0.22 μm-filtered solution in PBS (pH 7.4), 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 under sterile conditions after reconstitution.
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Performance Data
Evaluation of CAR expression
FACS Analysis of Anti-FMC63 CAR Expression
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 Alexa Fluor 647-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Cat. No. FM3-AM534) and negative control protein respectively (Fig. C and B), and non-transfected 293 cells were used as a control (Fig. A). Alexa 647 signal was used to evaluate the binding activity (QC tested).
FACS Analysis of Non-specific binding to PBMCs
5e5 of PBMCs were stained with Alexa Fluor 647-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Cat. No. FM3-AM534) and anti-CD3 antibody, washed and then analyzed with FACS. FITC signal was used to evaluate the expression of CD3+ T cells in PBMCs, and Alexa 647 signal was used to evaluate the non-specific binding activity to PBMCs (QC tested).
FACS Analysis of Non-specific binding to Human whole blood
Non-specificity of Alexa Fluor™ 647-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45) (Cat. No. FM3-AM534) 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 Alexa Fluor™ 647-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 Alexa Fluor™ 647 positive signals was used to evaluate the non-specific binding activity to human CD3+ cells.
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