Rabbit Anti-Human 2C6 scFv Monoclonal Antibody, Alexa Fluor 647 (2 Sizes)

The rabbit monoclonal antibody 1F5 specifically binds to the scFv region of a CD20-specific human monoclonal antibody (mAb, clone 2C6). CD20 (cluster of differentiate 20) is a protein that is expressed on the surface of B cells, starting at the pre-B cell stage and also on mature B cells in the bone marrow and in the periphery. CD20 is not expressed on hematopoietic stem cells, pro-B cells, or normal plasma cells. The scFv region of 2C6 has been used to develop CD20-specific chimeric antigen receptor (CAR) T cells utilized in clinical trials.

Shipping and Storage

• Store undiluted at 2-8°C.
• Avoid prolonged exposure to light.
• Avoid freeze/thaw cycle of the reagent.
• The monoclonal antibody was purified by Protein A.
• The antibody was conjugated with Alexa Fluor 647 under optimum conditions, and unincorporated dye was removed

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-Human 2C6 scFv Monoclonal 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-Human 2C6 scFv Monoclonal Antibody, 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.

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.