TotalSeq oligo-conjugated antibodies have been widely used to add protein data to single-cell RNA and DNA technologies. Now, the same trusted reagents, available in the new TotalSeq-Bn format, can be used to transform the 10x Genomics Visium CytAssist workflow into a multiomic spatial biology assay, with a simultaneous protein and RNA read-out from the same tissue sample.
TotalSeq-Bn antibodies are conjugated to an oligonucleotide containing a sequencing-compatible PCR handle, unique antibody barcode, and sequence complementary to the Visium CytAssist slide oligo pool.
Spatial Proteogenomics Workflow With 10x Genomics Visium CytAssist
The protein and gene expression libraries generated by the Visium CytAssist workflow are shown below. These libraries are analogous to the CITE-Seq libraries generated by the 10x Genomics Chromium platform for single-cell RNA and protein analysis. The main difference is that the 10x Genomics barcode unique for a single GEM is replaced by a spatial barcode that is unique for each of the thousands of spots in the Visium slide. Legacy TotalSeq-B antibodies are not compatible with Visium CytAssist.
Visium CytAssist Spatial Gene Expression – Probe-based Library
Visium CytAssist Spatial Protein Expression Library
TotalSeq™-Bn targets in the Visium Human Immune Cell Profiling Panel
The following TotalSeq™-Bn targets are available as part of the 10x Genomics Human Immune Cell Profiling Panel. Please contact 10x Genomics for information about the Panel and the Visium CytAssist assays.
Protein Marker | Description | Location | Clone | Isotype Control |
---|---|---|---|---|
Mouse IgG2a | Isotype control | Cell surface | MG2a-53 | N/A |
Mouse IgG1, κ | Isotype control | Cell surface | MOPC-21 | N/A |
Mouse IgG2b, κ | Isotype control | Cell surface | MPC-11 | N/A |
Rat IgG2a, κ | Isotype control | Cell surface | RTK2758 | N/A |
PanCK | Structural marker | Intracellular | AE-1/AE-3 | Mouse IgG1, κ |
PD1 | Checkpoint protein | Cell surface | NAT105 | Mouse IgG1, κ |
PCNA | Cell proliferation marker | Nuclear | PC10 | Mouse IgG2a |
BCL2 | Apoptotic cell marker | Mitochondrial outer membrane | 100 | Mouse IgG1 |
Vimentin | Mesenchymal cell marker | Intracellular | O91D3 | Mouse IgG2a |
CD8a | T cell marker | Cell surface | C8/144b | Mouse IgG1, κ |
CD45RA | Functional T cell marker | Cell surface | HI100 | Mouse IgG2b, κ |
CD45RO | Functional T cell marker | Cell surface | UCHL1 | Mouse IgG2a |
CD19 | B cell marker | Cell surface | R109 | Rabbit IgG |
PAX5 | B cell marker | Nuclear | 1H9 | Rat IgG2a, κ |
CD21 | Dendritic and B cell marker | Cell surface | BU32 | Mouse IgG1, κ |
CD68 | Macrophage marker | Cell surface | BL13756 | Mouse IgG1, κ |
CD163 | Macrophage marker | Cell surface | QA19A16 | Mouse IgG1, κ |
HLA-DR | Myeloid cell marker | Cell surface | LN3 | Mouse IgG2b, κ |
CD66b | Neutrophil marker | Cell surface | 6/40c | Mouse IgG1, κ |
CD138 | Plasma cell marker | Cell surface | DL-101 | Mouse IgG1, κ |
TotalSeq™-Bn Single Catalog Products
TotalSeq™-Bn single conjugate antibodies can be supplemented into 10x Genomics Human Immune Cell Profiling Panel. This offers the ability to expand the number of targets that can be interrogated by the Visium CytAssist Spatial Gene and Protein Expression assay. In order to optimize the concentration(s) and pool the additional antibody marker(s), refer to 10x Genomics Custom Add-on Antibody Optimization guide. For more information regarding the TotalSeq™-Bn single antibodies, please reach out to your local BioLegend Field Application Scientist or BioLegend’s Technical Service team.
Compatibility Information | |
---|---|
10x Genomics | Visium CytAssist Spatial Gene and Protein Solution with Capture Sequence technology (FFPE) and 10x Genomics Data Analysis Software (Space Ranger and Loupe Browser) |
Next-generation sequencing | Compatible with Illumina instruments |
Tissue | FFPE tissue |
Slide capture area | 6.5 mm2 or 11 mm2 |
The TotalSeq™-Bn format is not compatible with 10x Genomics Chromium single-cell solutions and CITE-Seq applications. More information about TotalSeq™ in single-cell applications and CITE-Seq can be found here.
Explore Additional Resources
Learn more about the Visium spatial solution:
- Visium Spatial Gene and Protein Expression
- Visium data analysis pipeline (Space Ranger)
- Visium data visualization (Loupe Browser)
- 10x Genomics Custom Add-on Antibody Optimization guide
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