scFv region of a CD19-specific mouse mAb clone FMC63
Aqueous buffered solution containing protein stabilizer and ≤0.03% sodium azide.
The mouse monoclonal antibody M19H specifically binds to the scFv region of a CD19-specific mouse monoclonal antibody (mAb, clone FMC63). CD19 antigen is a B-cell specific cell surface antigen, which is expressed in all B-cell lineage malignancies and normal B-cells. The scFv region of FMC63 has been used to develop CD19-specific chimeric antigen receptor (CAR) T cells utilized in clinical trials.
Flow cytometric analysis of anti-CD19-CAR expression on human T cell line Jurkat cells. Jurkat cells were lentivirally transduced with anti-CD19-CAR and cultured. 2×105 cells were stained for the expression of anti-CD19-CAR with Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Biotin antibody (Cat. No. 300414, right panel). Secondary staining was carried out with Streptavidin PE (Cat. No. 700032). Non-transduced Jurkat cells were used as a control for gating of CAR expression (left panel). Acquisition of >10, 000 events was performed.
Preparation and Storage
Shipped at 2-8°C. Store at 2-8°C short term (1-2 weeks). Store at -20°C in small aliquots for long term storage. Avoid freeze/thaw cycle.
The antibody was purified by Protein A.
The antibody was conjugated with biotin under optimum conditions.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
1. Harvest the cells and wash the cells once by FACS buffer (PBS containing 2% of BSA).
2. Count the cells number and the viability, aliquot up to 2×105 live cells into each tube. (Note: the cell viability must be ≥ 95%.)
3. Wash the cells once by FACS buffer.
4. Resuspend cells in 100 mL of diluted Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Biotin antibody (Cat. No. 300414, 1:100 diluted in FACS buffer) for 30 min at 4°C.
5. Wash the cells twice by FACS buffer.
6. Resuspend cells in 100 mL of diluted Streptavidin PE (Cat. No. 700032, 1:200 diluted in FACS buffer.) for 30 min at 4°C.
7. Wash the cells 3 times by FACS buffer and resuspend the cells in 200 mL PBS per sample.
8. Transfer the cells into flow tube and analyze on Flow Cytometer. Acquisition of >10, 000 events is performed.