Flow cytometry is an essential technique for immunology and cell biology for analyzing the properties of each cell in a sample population using fluorescence and light scattering.

Flow cytometry has broad applications including analysis of cell surface and intracellular markers, cell viability, apoptosis, cell cycle analysis, cell phenotyping and for cell sorting.

Most flow cytometry experiments involve the use of fluorescent proteins expressed by cells or fluorochrome-conjugated antibodies.

Proteintech offers a wide range of antibodies that are ideal for use in flow cytometry applications:

Many of Proteintech’s unconjugated antibodies can label extracellular and intracellular protein targets when paired with appropriate secondary antibodies. Below (Figure 1) are examples of extracellular cell adhesion molecule E-cadherin and intermediate filament Lamin proteins present in the nucleus of the cell:

Figure 1. Flow cytometry reagents for analyzing both extracellular and intracellular targets.

Figure 1. Left: HepG2 cells were stained with E-cadherin antibody (20874-1-AP, red) and IgG control antibody (blue) with anti-rabbit Alexa Fluor®488 secondary antibody. Right: HEK-293T cells were stained with Lamin A/C antibody (10298-1-AP, red) and IgG control antibody (blue) with anti-rabbit Alexa Fluor™ 488 secondary antibody.

Proteintech also offers primary antibodies directly conjugated to popular fluorophores that are compatible with practically any flow cytometer setup and do not require the use of secondary antibodies. These directly conjugated antibodies are perfect for multiplex flow cytometry experiments. Below (Figures 2 and 3) are examples of human blood samples stained with our anti-CD3 and anti-CD4 directly conjugated antibodies.

Figure 2. FITC-Anti-Human CD3 (FITC-65151, clone UCHT1) in flow cytometry

Figure 2.  0.1 mL human whole blood cells were surface stained with FITC-Anti-Human CD3 (FITC-65151, clone UCHT1). Samples were then treated with red blood cell lysis buffer and were gated for analysis of CD3 staining. Cells in the lymphocyte (red histogram), monocyte (green histogram), or granulocyte (blue histogram) gates (E1, E2, E3) were used to compare CD3 staining.

Figure 3. APC-Anti-Human CD4 (APC-65134, clone OKT4) in flow cytometry.

Figure 3. 1X10^6 human peripheral blood lymphocytes were surface stained with 0.20 ug APC-Anti-Human CD4 (APC-65134, clone OKT4) (red) or 0.20 ug APC-Mouse IgG2b isotype control (blue) and blank control (green). Samples were not fixed.