TotalSeq - Human TBNK Cocktail

The lyophilized cocktail provides convenience when processing several samples at the same time or at different time points, reducing inconsistencies associated with pipetting and handling multiple vials and samples. It also provides optimized and titrated antibodies that are ready to use and packaged in convenient single-use vials.

Recommended Usage

This panel has been optimized on human PBMCs. Using lysed whole blood additional TotalSeq™ antibodies may require further optimization. Please contact our technical service group for further information.

Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer`s intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer`s sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.Application Notes

Protocol for using TotalSeq™-A Human TBNK Panel

Antibody reconstitution

1. Bring up to room temperature for 5 minutes one vial of Human TotalSeq™-A TBNK panel.
2. Spin down at 10,000xg for 30 seconds at room temperature
3. Resuspend in 50 µL of Cell Staining Buffer (Cat. Nos. 420201) or similar, replace cap, vortex for 10 seconds.
   Note: Other buffer or solutions have not been tested
4. Incubate at room temperature for 5 minutes

Cell labeling

• Prepare human PBMCs following a suitable protocol.
• Resuspend cells at a concentration of 2.5x107 cells/mL in staining buffer.
• Make note of cell viability (>95%) and also include dead cells in the total cell count.
• If high cell death is observed, live cell enrichment (e.g. by Flow Cytometry) is recommended.

1. Aliquot 40 µL of cells (106 cells) into 12 x 75mm FACS tubes.
2. Add 5 µL of Human TruStain FcX™ (Fc Receptor Blocking Solution, Cat. Nos. 422301 & 422302), or 0.5 µL of TruStain FcX™ PLUS (anti-mouse CD16/32) Antibody (Cat. No. 156603 & 156604) diluted in 4.5 µL of staining buffer.
   Note: We no longer recommend the use of dextran during blocking/staining. If you have any questions please contact BioLegend Technical Support.
3. Incubate for 10 minutes at 4°C
4. Add the TotalSeq™ antibody panel. Use the entire content of the reconstituted tube.
5. Incubate 4°C for 30 minutes and proceed to Section I, step 7 of the TotalSeq™-A Protocol for 10x Single Cell Reagent Kit v3.1