This AbD Serotec Four-Color Human Flow Kit can be used for single-step identification of human helper/inducer naïve (CD3+CD4+CD45RA+CD62L+) and helper/inducer memory (CD3+CD4+CD45RA-CD62L+ or CD3+CD4+CD45RA-CD62L-) T-cell subsets.
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, ζ), consisting of two heterodimers (εγ, εδ) and one homodimer (ζζ). 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 20kDa CD3ε chain.
CD4 is a single chain transmembrane glycoprotein and member of the immunoglobulin superfamily, which interacts directly with class II molecules of the major histocompatibility complex (MHCII), and is the primary receptor for the human immunodeficiency virus (HIV). CD4 is expressed by the helper/inducer (CD3+CD4+) T-cell subset, the majority of thymocytes, and at low levels by monocytes and tissue macrophages. The CD4 lymphocyte count provides an excellent indication of immune system health. Clone RPA-T4 binds to the D1 domain (CDR1 and CDR3 epitopes) of the CD4 antigen.
CD45RA is an isoform of CD45, a member of the protein tyrosine phosphatase family, otherwise known as leucocyte common antigen (LCA), which acts as a positive regulator of T-cell co-activation, and is essential for the activation of T-cells through the antigen receptor. CD45RA is expressed by naïve T-cells, but not by activated and memory T-cells. Consequently, clone F8-11-13 can be used to distinguish between these cell populations.
CD62L is a cell surface adhesion protein, and member of the selectin/LECAM family, otherwise known as leukocyte-endothelial cell adhesion molecule 1 (LECAM1), which acts as a mediator of lymphocyte homing to high endothelial venules (HEVs), within peripheral lymphoid tissues. CD62L is constitutively expressed by all leucocytes including naïve T-cells, but only by certain subpopulations of memory T-cells. Levels of serum CD62L can be used as an indicator of inflammation.
Product Form
Four color combination consisting of FITC, RPE, RPE-Cy5.5 and APC conjugated monoclonal antibodies mixed in optimal ratio - lyophilised.
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 visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
Flow Cytometry
Neat
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Flow Cytometry
Use 10ul of the suggested working dilution to label 1x106cells in 100ul.
Storage
Prior to reconstitution store at +4oC.
After reconstitution store at +4oC.
DO NOT FREEZE.
This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
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