Watch How it Works


   

Walk Through the TotalSeq™ Workflow

 

1. Stain cells with TotalSeq™ reagents.

Titrate the antibodies to determine the ideal staining concentration for your sample. All TotalSeq™ reagents can be titrated by flow cytometry using a PE-conjugated format of the same clone.
2. Transfer labeled cells containing Antibody-Derived Tags (ADTs) onto a compatible sequencing platform.

 

This diagram illustrates the CITE-seq workflow using Drop-Seq and TotalSeq™-A reagents. Other compatible methods such as the Chromium platform from 10x Genomics may also be used. 


In step 2c, while TotalSeq™-B and –C reagents use a different capture sequence for hybridization, the overall workflow remains similar.

3. Amplify both cDNA and antibody-derived oligo libraries independently.

4. Pool libraries at the desired ratio for sequencing.

Improved Clustering of Distinct Cell Populations with TotalSeq™ Reagents



 

Clustering of approximately 5,000 CITE-seq single-cell expression profiles of PBMCs reveals distinct cell populations based on transcriptome analysis. The left panel shows a two-dimensional representation (tSNE) of global gene expression relationships among all cells. Major cell types in peripheral blood can be discerned based on marker gene expression as indicated. The right panels show mRNA (blue) and corresponding Antibody-Derived Tag (ADT, green) signal for the CITE-seq antibody panel projected on the tSNE plot. Darker shading corresponds to higher levels measured.