Description

The rabbit monoclonal antibody R19M 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. *

Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified by Protein A.
The antibody was conjugated with Alexa Fluor 647 under optimum conditions, and unincorporated dye was removed. *

Product Notices

• Since applications vary, each investigator should titrate the reagent to obtain optimal results.
• 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.

FACS Protocoal

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. Resuspend cells in 100 µL of diluted Anti-Mouse FMC63 scFv mAb (R19M), Alexa Fluor 647 (Cat. No. 200101, 1:100 diluted in FACS buffer) for 30 min at 4°C.
4. Wash the cells 3 times by FACS buffer and resuspend the cells in 200 µL PBS per sample.
5. Transfer the cells into flow tube and analyze on Flow Cytometer. Acquisition of >10, 000 events is performed.