Description

The rabbit polyclonal antibody R19 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.

Item Information
Catalog #	Size/concentration	Price	Antibody Registry Number
500014	100 tests	$400	AB_2857948

Host species	Rabbit
Antibody type	Polyclonal
Immunogen	scFv region of a CD19-specific mouse mAb clone FMC63
Reactivity	Mouse
Application	FCM
Storage buffer	Aqueous buffered solution containing protein stabilizer and ≤0.03% sodium azide

Preparation and Storage

Shipped at 4°C. Store at 4°C short term (1-2 weeks). Store at -20°C in small aliquots for long term storage. Avoid freeze/thaw cycle.
The polyclonal antibody was purified by Protein A.
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Product Notices

• Since applications vary, each investigator should titrate the reagent to obtain optimal results.
• An isotype control should be used at the same concentration as the antibody of interest.
• 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 Protocol

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 Blocking Reagent (Cat. No. 700000, 1:100 diluted in FACS buffer) for 45 min at 4°C.
4. Wash the cells once by FACS buffer.
5. Resuspend cells in 100 µL of diluted Biotin Rabbit Anti-Mouse FMC63 scFv antibody (Cat. No. 500014, 1:100 diluted in FACS buffer) for 30 min at 4°C.
6. Wash the cells once by FACS buffer.
7. Resuspend cells in 100 µL of diluted StreptavidinPE (Cat. No. 700006, 1:200 diluted in FACS buffer) for 30 min at 4°C.
8. Wash the cells 3 times by FACS buffer and resuspend the cells in 200 µL PBS per sample.
9. Transfer the cells into flow tube and analyze on Flow Cytometer. Acquisition of >10, 000 events is performed.

Technical Data Sheet