$530.00

The rabbit monoclonal antibody R19M 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 #  200102 
Size/Concentration  100 tests 
Price  $1,925.00 
Antibody Registration Number  AB_2857946 

Specification 
Host Species  Rabbit 
Antibody Type   Monoclonal 
Antigen Purification  Protein A 
Fluorescent Dye  Alexa Fluor 647 
Excitation (nm)  650 
Reactivity  Mouse 
Immunogen  scFv region of a CD19-specific mouse mAb clone FMC63 
Storage Buffer  Aqueous buffered solution containing protein stabilizer and ≤0.03% sodium azide 
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 

  • Reagents Required
    • Phosphate Buffered Saline (PBS with a pH ~7.4). 
    • FACS Buffer (PBS containing 2% of BSA). 
    • Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647
  • Preparation of Reagents
    1. Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647
      • The antibody must be diluted before use for FACS.
      • Dilute the antibody 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 Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647 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

  • Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  • 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.

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..

Flow Cytometry FACS Protocol 

  • Reagents Required
    • Phosphate Buffered Saline (PBS with a pH ~7.4). 
    • FACS Buffer (PBS containing 2% of BSA). 
    • Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647
  • Preparation of Reagents
    1. Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647
      • The antibody must be diluted before use for FACS.
      • Dilute the antibody 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 Rabbit Anti-Mouse FMC63 scFv Monoclonal Antibody (R19M), Alexa Fluor 647 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.

Citations

  • Agarwal S, Aznar MA, Rech AJ, Good CR, Kuramitsu S, Da T, Gohil M, Chen L, Hong SA, Ravikumar P, Rennels AK, Salas-Mckee J, Kong W, Ruella M, Davis MM, Plesa G, Fraietta JA, Porter DL, Young RM, June CH. Deletion of the inhibitory co-receptor CTLA-4 enhances and invigorates chimeric antigen receptor T cells. Immunity. 2023 Oct 10;56(10):2388-2407.e9. doi: 10.1016/j.immuni.2023.09.001. Epub 2023 Sep 29. PMID: 37776850; PMCID: PMC10591801.
  • Cheng, Jiali, et al. Monitoring Anti‐CD19 Chimeric Antigen Receptor T Cell Population by Flow Cytometry and Its Consistency with Digital Droplet Polymerase Chain Reaction. Aug. 2022, https://doi.org/10.1002/cyto.a.24676.
  • Cheng J, Yan J, Liu Y, Shi J, Wang H, Zhou H, Zhou Y, Zhang T, Zhao L, Meng X, Gong H, Zhang X, Zhu H, Jiang P. Cancer-cell-derived fumarate suppresses the anti-tumor capacity of CD8+ T cells in the tumor microenvironment. Cell Metab. 2023 Jun 6;35(6):961-978.e10. doi: 10.1016/j.cmet.2023.04.017. Epub 2023 May 12. PMID: 37178684.
  • Cheng J, Mao X, Chen C, Long X, Chen L, Zhou J, Zhu L. Monitoring anti-CD19 chimeric antigen receptor T cell population by flow cytometry and its consistency with digital droplet polymerase chain reaction. Cytometry A. 2023 Jan;103(1):16-26. doi: 10.1002/cyto.a.24676. Epub 2022 Aug 5. PMID: 35875964; PMCID: PMC10087589.
  • Lei W, Zhao A, Liu H, Yang C, Wei C, Guo S, Chen Z, Guo Q, Li L, Zhao M, Wu G, Ouyang G, Liu M, Zhang J, Gao J, Qian W. Safety and feasibility of anti-CD19 CAR T cells expressing inducible IL-7 and CCL19 in patients with relapsed or refractory large B-cell lymphoma. Cell Discov. 2024 Jan 9;10(1):5. doi: 10.1038/s41421-023-00625-0. PMID: 38191529; PMCID: PMC10774422.
  • Wu W, Zhou Q, Masubuchi T, Shi X, Li H, Xu X, Huang M, Meng L, He X, Zhu H, Gao S, Zhang N, Jing R, Sun J, Wang H, Hui E, Wong CC, Xu C. Multiple Signaling Roles of CD3ε and Its Application in CAR-T Cell Therapy. Cell. 2020 Aug 20;182(4):855-871.e23. doi: 10.1016/j.cell.2020.07.018. Epub 2020 Jul 29. PMID: 32730808.

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