Rabbit Anti-Mouse C11D5.3 scFv Polyclonal Antibody, Alexa Fluor 647 (100 Tests)

The rabbit polyclonal antibody specifically binds to the scFv region of a B-cell maturation antigen (BCMA) specific mouse monoclonal antibody (mAb, clone C11D5.3). BCMA is a protein that has been reported to be selectively expressed by B-lineage cells including multiple myeloma cells1 and restrictively expressed in both normal and malignant plasma cells at high levels2. The scFv region of C11D5.3 has been used to develop BCMA-specific chimeric antigen receptor (CAR) T cells utilized in clinical trials.

Shipping and Storage 

FACS Protocol
BioSwan reagents can be used with or without an isotype control to assess the amount of nonspecific antibody binding.
(Optional) For Whole Blood Sample

1. Pipette 1 μL Rabbit Anti-Mouse C11D5.3 scFv Polyclonal Antibody, 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 Rabbit Anti-Mouse C11D5.3 scFv Polyclonal Antibody, Alexa Fluor 647, dead cell staining solution and additional fluorochrome. Mix
5. gently and thoroughly.
6. Incubate for 25 minutes in the dark at room temperature (18-25°C).
7. 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.
8. Repeat step 6 twice.
9. Add a suitable amount of FACS buffer to resuspend cell and analysis by flow cytometry.

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

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 (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; or (b) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. 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.
2. 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.

References

1. Robert O. Carpenter et al., “B-Cell Maturation Antigen Is a Promising Target for Adoptive T-Cell Therapy of Multiple Myeloma,” Clinical Cancer Research 19, no. 8 (April 15, 2013): 2048–60, https://doi.org/10.1158/1078-0432.CCR-12-2422.
2. Bo Yu, et al., “BCMA-Targeted Immunotherapy for Multiple Myeloma,” Journal of Hematology & Oncology 13, no. 1 (December 2020): 125, https://doi.org/10.1186/s13045-020-00962-7.