$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 #  300413 
Size/Concentration  25 tests 
Price  $900.00 

Specification 
Host Species  Mouse 
Antibody Type   Monoclonal 
Antigen Purification  Protein A 
Fluorescent Dye  R-PE 
Excitation (nm)  565 
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 
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 4°C 
  • Avoid prolonged exposure to light 
  • Avoid freezing the reagent  

Flow Cytometry FACS Protocol

  • 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, Biotin (100 Tests) 
      • Streptavidin PE  
    • Preparation of Reagents 
      • Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Biotin (100 Tests) 
        • Protein must be diluted before use for FACS. 
        • Dilute the protein at a dilution of 1:100 in FACS buffer. 
      • Streptavidin PE 
      • Dilute streptavidin PE at a dilution of 1:200 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, Biotin antibody for 30 min at 4°C. 
      4. Wash the mixture of cells and antibody twice by centrifuge using FACS buffer. 
      5. Resuspend cells in 100 µL of diluted Streptavidin PE for 30 min at 4°C. 
      6. Resuspend the stained cells in 200 µL PBS per cell sample.  
      7. 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

For Research Use Only. Not for use in diagnostic or therapeutic procedures

  1. This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer and the buyer must not use this product or its components:
    for:

    1. diagnostic, therapeutic or prophylactic purposes; or
    2. manufacturing or quality assurance or quality control.
    3. and/or to sell or transfer this product or its components for resale, whether or not resold for use in research.
  2. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
  3. 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.
  4. 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.
  5. Any use of this product other than the permitted use without the express written authorization of BioSwan Company is strictly prohibited.
    Not for resales. BioSwan, the BioSwan Logo and all other trademarks are property of BioSwan Laboratories, Co., Ltd..

Following this protocol should enable the identification and analysis of cell populations expressing the FMC63 scFv marker.

Flow Cytometry 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, Biotin-conjugated (for 100 tests).
  • Streptavidin Phycoerythrin (PE) conjugate.

Preparation of Reagents

  1. Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Biotin-conjugated

    • Dilute the antibody to a 1:100 ratio in FACS buffer before use in flow cytometry assays.

  2. Streptavidin PE Conjugate

    • Prepare a 1:200 dilution of Streptavidin PE in FACS buffer.

Staining Protocol

  1. Cell Preparation

    • Harvest cells from their culture or source and wash once with FACS buffer.
    • Perform a cell count and assess viability. Ensure a viability of ≥95% with at least 2×10^5 viable cells.

  2. Primary Antibody Incubation

    • Resuspend the cells in 100 µL of the diluted Mouse Anti-Mouse FMC63 scFv Monoclonal Antibody, Biotin.
    • Incubate for 30 minutes at 4°C.

  3. Washing

    • After incubation, wash the cell-antibody mixture twice by centrifugation with FACS buffer to remove excess, unbound antibody.

  4. Secondary Antibody Incubation

    • Resuspend the cells in 100 µL of the diluted Streptavidin PE.
    • Incubate for another 30 minutes at 4°C.

  5. Final Resuspension

    • Resuspend the stained cells in 200 µL of PBS for each sample.

  6. Flow Cytometry Analysis

    • Transfer the stained cells to appropriate tubes for flow cytometry.
    • Analyze on the flow cytometer with a recommended acquisition rate of over 10,000 events.

Notes

  • Adjustments to dilutions, incubation times, or temperatures may be necessary based on specific experimental conditions or desired outcomes.
  • It’s crucial to maintain cells in a viable state throughout the process to ensure accurate analysis.

Citation List

  • An, J., Zhang, CP., Qiu, HY. et al. Enhancement of the viability of T cells electroporated with DNA via osmotic dampening of the DNA-sensing cGAS–STING pathway. Nat. Biomed. Eng 8, 149–164 (2024). https://doi.org/10.1038/s41551-023-01073-7
  • Sun M, Xu P, Wang E, Zhou M, Xu T, Wang J, Wang Q, Wang B, Lu K, Wang C, Chen B. Novel two-chain structure utilizing KIRS2/DAP12 domain improves the safety and efficacy of CAR-T cells in adults with r/r B-ALL. Mol Ther Oncolytics. 2021 Aug 28;23:96-106. doi: 10.1016/j.omto.2021.08.014. PMID: 34703879; PMCID: PMC8517091.
  • Wang Y, Zhong K, Ke J, Chen X, Chen Y, Shu W, Chen C, Hu S, Sun X, Huang H, Luo C, Liu L, Yang J, Zhang Y, Zhi H. Combined 4-1BB and ICOS co-stimulation improves anti-tumor efficacy and persistence of dual anti-CD19/CD20 chimeric antigen receptor T cells. Cytotherapy. 2021 Aug;23(8):715-723. doi: 10.1016/j.jcyt.2021.02.117. PMID: 33863641.
  • Yang L, Yin J, Wu J, Qiao L, Zhao EM, Cai F, Ye H. Engineering genetic devices for in vivo control of therapeutic T cell activity triggered by the dietary molecule resveratrol. Proc Natl Acad Sci U S A. 2021 Aug 24;118(34):e2106612118. doi: 10.1073/pnas.2106612118. PMID: 34404729; PMCID: PMC8403971.

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