Cell Analysis and Reagents

Multicolor products

The Custom Technology Team works with clients to make both qualitative and quantitative multicolor antibody cocktails for flow cytometry–based immunophenotyping.

These products include conjugated antibodies, compensation controls, buffers, and protocol guidelines. Optimized multicolor flow cytometry cocktails can streamline sample preparation, acquisition, and analysis, and improve standardization between experiments.

Intracellular proteomics (BD Phosflow)

CTT offers assay optimization, sample processing, and data delivery for BD™ Phosflow analysis. An intracellular phosphoprofiling tool, BD Phosflow technology allows scientists to examine cellular subpopulations in complex samples such as primary cells, whole blood, or peripheral blood mononuclear cells (PBMCs). Unlike traditional methods such as Western blotting, BD Phosflow enables researchers to analyze phosphoprotein signaling in single cells through the use of multiple cell surface markers. Using BD Phosflow alone, researchers can uncover rare cell subtypes with different signaling mechanisms.

Cell surface marker profiling

For researchers using stem cells in drug discovery or cell therapy, CTT offers cell surface marker profiling to identify cell state variations indicative of donor-to-donor variations, cell type specific markers, or treatment-specific changes. The team leverages the novel proteomics tool BD FACS™ CAP (Combinatorial Antibody Profiling), a flow cytometry–based method that uses a broad screening panel of more than 200 high-quality fluorescently-labeled BD antibodies.


Immunophenotyping of regulatory T cells

Immunophenotyping of regulatory T cells

The data shown demonstrates the characterization of regulatory T cells through multicolor flow cytometry. Human PBMCs were surface stained for CD4 and CD25, fixed, permeabilized, and stained intracellularly for FoxP3. The data shown were derived from an acquisition of 50,000 events in a lymphocyte gate, followed by CD4+ gating by fluorescence. A compound gating strategy by morphology, then side scatter vs. fluorescence, was used to identify FoxP3+ Treg cells shown in a final plot representing CD25 vs. FoxP3.

Cell surface marker profiling

Cell surface marker profiling

Graphical representation of BD FACS CAP screen showing relative expression of cell surface markers on multiple cell types from multiple donors. Color key represents percent positive cells.

Multiple phosphoproteins in different cell types studied by BD Phosflow

Multiple phosphoproteins in different cell types studied by BD Phosflow

This figure illustrates how BD Phosflow makes it possible to conduct the simultaneous analysis of multiple phosphoproteins in complex cell mixtures. In this experiment, BD Phosflow was used to analyze the effect of four different stimuli on cell signaling pathways in mouse splenic cell subsets (B cells, T cells, and CD11bhighcells). Differences in signaling pathway responses were uncovered between mouse splenocyte cultures stimulated in vitro and splenic cells stimulated in vivo, underscoring the importance of conducting studies in close to native conditions. Data were analyzed in Cytobank software (http://cytobank.org). Histograms are colored according to fold change in phosphorylated protein relative to unstimulated samples.

Data courtesy of Dr. Peter Krutzik and Dr. Matt Hale, Stanford University.


For Research Use Only. Not for use in diagnostic or therapeutic procedures.