Alexa Fluor™ 647-Labeled Monoclonal Anti-FMC63 Antibody, Mouse IgG1 (Y45)Star Staining

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.