Schematic diagram of Human Treg Panel Kit.
Product Details
Product Details
Product Specification
| Application | FCM |
| Reactivity | Hu |
| Positive Sample | Human PBMC |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 1% BSA, 0.09% sodium azide |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8 °C as supplied. |
Background
This four‑color panel is designed for detecting regulatory T cells (Tregs) in human peripheral blood or other biological samples. CD3 was included as a general T‑cell marker, and CD4 was included to identify helper T cells (CD4+), within which Tregs are predominantly found. CD25 is a marker often used to detect Tregs. Finally, CD127 or FOXP3 was included to confirm the Treg population, as Tregs typically exhibit a CD25+CD127− phenotype or a CD25+FOXP3+ phenotype; it is important to note that FOXP3 is an intracellular marker and requires fixation and permeabilization. The panel is compatible with most three‑laser flow cytometers. If desired, the panel can serve as a back bone panel, allowing for the addition of antibodies conjugated to various fluorophores to detect other markers of interest.
Picture
Picture
FC
Human PBMCs were stained using the Human Treg Panel Kit consisting of anti- Human CD3 Alexa Fluor® 488, anti- Human CD4 PerCP-Cy5.5, anti- Human CD25 PE and anti- Human CD127 APC conjugated antibodies.
To analyze the data:
1. Create an FSC-A vs SSC-A dot plot and draw a gate around the lymphocyte population.
2. Create a Fixable Viability Dye 545 (S0B88802) vs SSC-A dot plot with the leukocyte gate applied. Draw a gate around the live cell population.
3. Create a CD3 Alexa Fluor® 488-A vs SSC-A dot plot show the T cells with the live cell gate applied.
4. Create a CD4 PerCP-Cy5.5-A vs SSC-A dot plot with the CD3+ gate applied.
5. Create a CD25 PE -A vs CD127 APC-A dot plot and apply the CD4+ gate. Create a gate to encompass CD25+ CD127- cells (Treg cells).
