Intracellular CITE-seq Assay Using BD^® AbSeq Antibody-Oligos

Expand the power of CITE-seq to intracellular proteomics with BD^®AbSeq Antibody-Oligos and associated reagents

Redefine CITE-seq

Profile intracellular proteins, surface proteins and RNA in one single-cell experiment

The Intracellular CITE-seq Assay using BD^® AbSeq Antibody-Oligos:

Enables simultaneous RNA, surface protein and intracellular protein detection
Supports high-plex proteomics profiling (~100 protein markers) at the single-cell level
Allows a safe stopping point during the workflow without compromising assay sensitivity
Works with the BD^® Single-Cell Multiplexing Kit to enable sample multiplexing
Drives deeper understanding of cellular functions such as signal transduction dynamics and transcriptional regulation

Get more information from the Intracellular CITE-seq Assay using BD^® AbSeq Antibody-Oligos brochure.

List of Available Intracellular BD^® AbSeq Ab-Oligo Specificities

Specificity	Clone
Caspase-3	C92-605
PARP (Cleaved)	F21-852
H2AX (pS139)	N1-431
T-bet	O4-46
Granzyme B	GB11
Helios	22F6
BCL-6	K112-91
GATA3	L50-823
Ki-67	B56
Stat6 (pY641)	18/P-Stat6
p38 MAPK (pT180/pY182)	36/p38 (pT180/pY182)
Chromogranin A	S21-537
Sox2	O30-678
Sox17	P7-969

Applications

Intracellular BD^® AbSeq Antibody-Oligos can be used to characterize a heterogeneous subset of T follicular helper (T_FH) cells in human tonsil

A combination of intracellular CITE-seq, surface CITE-seq and scRNA-seq was used to deepen our understanding of the differentiation states and subsets of cells present at low abundance, such as T_FH cells.

We characterized the heterogeneous subsets of T_FH and circulating T_FH (cT_FH) cells using a combination of intracellular CITE-seq, surface CITE-seq and whole transcriptome single-cell RNA sequencing (scRNA-seq) analysis. Germinal center and cT_FH cells were enriched by sorting memory CD4⁺, CXCR5⁺, PD-1⁺ from a tonsil (n = 1) and memory CD4⁺ CXCR5⁺ cells from a matched blood sample and a healthy blood donor (n = 2). We examined the expression of Bcl-6 and other transcription factors, using protein and RNA expression data, across pre-T_FH (an early development stage in the germinal center), T_FH and cT_FH cells.

Tonsillar T_FH cells were enriched and stained with both surface and intracellular BD^®AbSeq Antibody-Oligos.

A) Annotated UMAP of tonsil, colored by cell type. B) Statistically significant (Bonferroni adjusted P value (P val adj) > 0.05) versus fold change for differentially expressed genes (DEG) between T_FH cells compared to pre-T_FH cells. C) Heatmap visualizing the gene signatures and surface and intracellular BD^®AbSeq Antibodies used to identify cell subsets within the tonsil. D) Top panels display surface and intracellular BD^®AbSeq Antibodies used to identify T_FH cell (left), T regulatory cells (T_Reg) (middle) and B cells (right). Bottom panels display corresponding RNA signatures. E) Surface and intracellular BD^® AbSeq Antibodies correlate with RNA expression within the tonsil.