$240.00

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.

Item Information 
Catalog #  300406 
Size/Concentration  100 Tests 
Price  $900.00 

Specification 
Host Species  Mouse 
Antibody Type   Monoclonal 
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  R-PE 
Excitation (nm)  565 
Application  FCM 

Shipping and Storage 

Shipping   The product is shipped at 2-8° C.  
Stability & Storage   To provide optimal stability of reagent:   

  • Store undiluted at 2-8° C
  • Avoid prolonged exposure to light.
  • The monoclonal antibody was purified by Protein A
  • The antibody was conjugated with PE under optimum conditions, and unincorporated dye was removed.

FACS Protocol 

  • Reagents Required 
    • Phosphate Buffered Saline (PBS with a pH ~7.4).  
    • FACS Buffer (PBS containing 2% of BSA).  
    • Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, PE (100 Tests) 
  • Preparation of Reagents 
    • Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, PE (100 Tests) 
      • Protein must be diluted before use for FACS. 
      • Dilute the protein at a dilution of 1:100 in FACS buffer. 
  • Staining Protocol 
    1. Harvest the cells from source and wash the cells once using FACS buffer. 
    2. Perform a cell count and a cell viability count. Cell viability must be ≥ 95% with a cell number above 2×105 live cells. 
    3. In a round-bottom test tube, resuspend cells in 100 µL of diluted Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, PE for 30 min at 4°C. 
    4. Wash the mixture of cells and antibody 3 times by centrifuge using FACS buffer. 
    5. Resuspend the stained cells in 200 µL PBS per cell sample.  
    6. Transfer the stained cell into a desired flow tube for analyzation on desired Flow Cytometer. The recommended acquisition rate is >10,000 events. 

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Antibody solutions containing ProClin 300 should be handled with care. Do not take internally and avoid all contact with the skin, mucosa
    and eyes.

Intellectual Product Notices

  • Conditions: The information disclosed herein is not to be construed as a recommendation to use the above product in violation of any patents. BioSwan will not be held responsible for patent infringement or other violations that may occur with the use of our products. Purchase does not include or carry any right to resell or transfer this product either as a stand-alone product or as a component of another product. Any use of this product other than the permitted use without the express written authorization of BioSwan Company is strictly prohibited. For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resales. BioSwan, the BioSwan Logo and all other trademarks are property of BioSwan Laboratories, Co., Ltd.

FACS Analysis Protocol

Reagents Required

  • Phosphate Buffered Saline (PBS, pH ~7.4).
  • FACS Buffer: PBS supplemented with 2% Bovine Serum Albumin (BSA).
  • Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Phycoerythrin (PE) conjugated (for 100 tests).

Preparation of Reagents

  1. Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, PE-Conjugated
    • This antibody needs to be diluted before use in Fluorescence-Activated Cell Sorting (FACS) applications.
    • Prepare the antibody solution by diluting at a ratio of 1:100 in FACS buffer.

Staining Protocol

  1. Cell Harvesting and Washing
    • Harvest cells from the culture or specimen source, followed by a single wash with FACS buffer.
  2. Cell Counting and Viability Assessment
    • Perform a cell count and determine cell viability. Ensure a viability of ≥95% and a total cell count exceeding 2×10^5 viable cells.
  3. Primary Antibody Staining
    • Resuspend the harvested cells in 100 µL of the diluted Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, PE conjugate.
    • Incubate for 30 minutes at 4°C in a round-bottom test tube.
  4. Washing
    • After incubation, wash the cell and antibody mixture three times by centrifugation with FACS buffer to remove unbound antibody.
  5. Resuspension
    • Following the final wash, resuspend the stained cells in 200 µL of PBS for each sample.
  6. Flow Cytometry Analysis
    • Transfer the stained cells into appropriate flow cytometry tubes.
    • Analyze on a flow cytometer with a recommended acquisition rate of over 10,000 events.

Notes

  • Light sensitivity: As PE is a fluorescent dye, keep samples protected from light post-staining to prevent photobleaching.
  • If nonspecific binding is a concern, consider including a control sample stained with an isotype-matched, non-specific PE-conjugated antibody.
  • Adjustments to antibody dilutions or incubation times may be necessary based on specific experimental conditions, antibody performance, or cell types.

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