Aqueous buffered solution containing protein stabilizer and ≤0.03% sodium azide
1A3 specifically binds to the scFv region of a B-cell maturation antigen (BCMA) specific mouse monoclonal antibody (mAb, clone C11D5.3). B-cell maturation antigen (BCMA) is a protein that has been reported to be selectively expressed by B-lineage cells including multiple myeloma cells and plasma cells. The scFv region of C11D5.3 has been used to develop BCMA-specific chimeric antigen receptor (CAR) T cells utilized in clinical trials.
Flow cytometric analysis of anti-BCMA CAR expression on human cell line Jurkat cells. Jurkat cells were lentivirally transduced with anti-BCMA CAR and cultured. 2×105 cells were stained for the expression of anti-BCMA CAR with Rabbit Anti-Mouse C11D5.3 scFv Monoclonal Antibody, Alexa Fluor 647 (Cat. No. 200702, right panel). Non-transduced Jurkat cells were used as a control for gating of CAR expression (left panel).
Preparation and Storage
Shipped at 2-8°C. Store at 2-8°C. Avoid freeze/thaw cycle.
The monoclonal antibody was purified by Protein A.
The antibody was conjugated with Alexa Fluor 647 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.
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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 Rabbit Anti-Mouse C11D5.3 scFv Monoclonal Antibody, Alexa Fluor 647 (Cat. No. 200702, 1:100 diluted in FACS buffer) for 30 min at 4°C.
5. Wash the cells 3 times by FACS buffer and resuspend the cells in 200 mL PBS per sample.
6. Transfer the cells into flow tube and analyze on Flow Cytometer. Acquisition of >10, 000 events is performed.