Four-Color Human CD3/CD16+CD56/CD45/CD19 Flow Kit can be used for single-step identification of T-cells (CD3+), B-cells (CD3-CD19+) and NK cells (CD3-CD16+ and/or CD56+), useful in the study of NK-mediated cytotoxicity against malignant and virally infected cells and the characterization and monitoring of immunodeficiency and autoimmune diseases. CD45 is a 180-240 kDa complex glycoprotein and member of the protein-tyrosine phosphatase family, otherwise known as leucocyte common antigen (LCA), and is expressed on all haematopoietic cells with the exception of mature erythrocytes and platelets. CD45 is a complex molecule existing in a number of isoforms including CD45RA and CD45RO. Clone F10-89-4 reacts with all isoforms. CD3 is a member of the immunoglobulin superfamily, which acts as a mediator of signal transduction, through association with the α/β or γ/δ T-cell receptor (TCR). Mammalian CD3 is a multimeric protein composed of four distinct polypeptide chains (ε, γ, δ, θ), consisting of two heterodimers (εγ, εδ) and one homodimer (θθ). CD3 is expressed by a high-percentage of circulating peripheral T-cells and is considered a pan T-cell marker. Clone UCHT1 specifically recognizes the ~20 kDa CD3ε chain. CD19 is a member of the immunoglobulin super family, which acts as a critical signal transduction molecule and regulator of B-cell development, differentiation and activation. CD19 is expressed on follicular dendritic cells and B-cells, from earliest recognizable B-lineage cells to B-cell blasts, but is lost on maturation to plasma cells. As such, CD19 is a commonly used pan B-cell marker. CD16/CD56, used in combination, facilitate the identification of natural killer (NK) cell sub-populations which express CD16+/CD56+, CD16+/CD56- or CD16-/CD56+ but lack CD3 and T-cell receptor (TCR) expression. There are important functional differences between these NK cell subsets. CD16-/CD56+(bright) cells, a minority in peripheral blood but representing the majority of NK cells in secondary lymphoid tissues, exhibit a high cytokine production capacity but are less effective at mediating antibody-dependent cellular cytotoxicity (ADCC) and natural NK cytotoxicity than other subsets.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

Application Name	Verified	Min Dilution	Max Dilution
Flow Cytometry			Neat

Flow Cytometry

Use 10ul of the suggested working dilution to label 1x10⁶ cells in 100ul.