Lymphocytes were stained according to standard protocols. Briefly, for staining surface markers, cells were resuspended in fluorescence‐activated cell‐sorting (FACS) buffer at a concentration of 2×106 cells/mL. Nonspecific antibody binding was blocked with anti–rat CD32 antibody (BD Biosciences, Franklin Lakes, NJ) followed by staining with specific antibodies (1:1000, all from BioLegend, San Diego, CA) for 60 minutes on ice in the dark. Antibodies used for flow cytometry included FITC‐CD3, APC‐CD3, PECy7‐CD4, PE‐CD25, FITC‐CD8, APC‐CD45RA, and Alexa Fluor 647‐Foxp3. Intracellular Foxp3 staining was performed after cell surface staining using the True Nuclear transcription factor staining kit (BioLegend) according to the manufacturer's protocol. Stained cells were washed twice and resuspended in FACS buffer and analyzed or sorted on a BD FACSAria II flow cytometer (BD Biosciences).
Antirat cd32 antibody
Manufactured by BD
The Antirat CD32 antibody is a laboratory reagent used in scientific research. It is designed to specifically recognize and bind to the CD32 protein found on the surface of certain cells in rats. The CD32 protein plays a role in immune system function. The Antirat CD32 antibody can be used to identify and study cells expressing this protein.
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Lab products found in correlation
3 protocols using antirat cd32 antibody
1
Lymphocyte Immunophenotyping by Flow Cytometry
2
Alloreactive T-cell identification
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Donor thymocytes blocked with antirat CD32 antibody (D34-485, BD Biosciences, San Jose, CA) were incubated with the serum samples of recipients. After washing, the cell samples were incubated with fluorescein isothiocyanate-conjugated mouse polyclonal antirat IgG antibody (Invitrogen) and phycoerythrin-conjugated mouse antirat CD3 antibody (eBioG4.18, Invitrogen). Samples were analyzed by flow cytometry (FACSCanto II, BD Biosciences).
3
Quantification of Regulatory T Cells
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The Fc receptor was blocked by incubating the cells with antirat CD32 antibody (BD Biosciences, San Jose, CA) for 30 min. The cells were then surface stained using anti-rat CD4-APC and anti-rat CD25-PE antibodies from Biolegend (San Diego, CA). After treating with antibodies for 40 min, cells were washed and stained with FITC labeled FoxP3 antibody (eBiosciences, San Diego, CA) using FoxP3 staining kit according to manufacturer's instructions (ebiosciences). The stained cells were analyzed using FACS Calibur (BD Biosciences). The percentage of CD4 + CD25 + FoxP3 + Tregs was calculated using WinMDI [18, (link)19] (link).
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