Ra1D6 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 #Â | 200801Â |
| Size/Concentration | 25 tests |
| Price | $530.00 |
|
Specification |
|
| Antibody Type  | Primary antibody, Recombinant antibody |
| Clone  | Ra1D6 |
| Antigen Purification | Protein A |
| Reactivity | Mouse |
| Immunogen | scFv region of a CD19-specific mouse mAb clone FMC63 |
| Storage Buffer | Aqueous buffered solution containing protein stabilizer and ≤0.05% ProClin 300. |
| Fluorescent Dye | Alexa Fluor 647 |
| Excitation (nm)Â | 650Â |
| Application | FCM |
Shipping and Storage
| Shipping  | The product is shipped at 2-8°C.  |
| Stability & Storage  | To provide optimal stability of reagent:  Â
|
FACS Protocol
- Â If you want to set a control, the recommended reagent to use is the following: Rabbit IgG Isotype Control, Alexa Fluor 647 (Product No. 700302).
(Optional) For Whole Blood Sample
1. Pipette 1 μL Anti-Mouse FMC63 scFv Monoclonal Antibody (Ra1D6), hIgGC, Alexa Fluor 647 into the bottom of the tube.
2. Add dead cell staining solution and additional fluorochrome conjugated antibodies into the bottom of the tube.
3. Pipette 100 µL of well-mixed, anticoagulated whole blood into the bottom of the tube. Mix gently and thoroughly.
Note Avoid smearing sample down the side of the tube. If the sample remains on the side of the tube, it will not be stained with the reagents.
4. Incubate for 25 minutes in the dark at room temperature (18-25°C).
5. Pipette Red Blood Cell Lysis Solution to the tube. Mix gently and thoroughly. Incubate for 15 minutes in the dark at room temperature (18-25°C).
6. Add 500 μL FACS buffer to the tube. Mix well and centrifuge at 300g for 5 minutes at room temperature (18-25°C). Aspirate supernatant
completely.
7. Repeat step 6 twice.
8. Add a suitable amount of FACS buffer to resuspend cell and analysis by flow cytometry.
(Optional) For Cell Sample
1. Harvest the cells and wash the cells twice by FACS buffer.
2. Count the cells number and the viability.
3. Resuspend the cell suspension to a concentration up to 1×106 nucleated cells per 100 μL of buffer.
4. Add 1 μL Anti-Mouse FMC63 scFv Monoclonal Antibody (Ra1D6), hIgGC, Alexa Fluor 647, dead cell staining solution and additional fluorochrome.
Mix gently and thoroughly
5. Incubate for 25 minutes in the dark at room temperature (18-25°C).
6. Add 500 μL FACS buffer to the tube. Mix well and centrifuge at 300 g for 5 minutes at room temperature (18-25°C). Aspirate supernatant
completely.
7. Repeat step 6 twice.
8. Add a suitable amount of FACS buffer to resuspend cell and analysis by flow cytometry
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