Recipient tumor-bearing mice were sacrificed 8 h after injection of DiR-labeled CTLs. The tumors of these mice were isolated. Single cell suspensions of tumor tissues were prepared and the CTL content was detected using flow cytometry. The single cell suspension was adjusted to 1–2×107 cells/ml and 1 ml was inserted into a 1.5-ml Eppendorf tube. Following washing two times with PBS, the cells were suspended in 5 µl CD3 antibodies (BD Biosciences; cat. no. 6203-25, 1:20 dilution) respectively, and incubated in the dark at 4°C for 30 min. Subsequently, cells were washed with PBS two times again and suspended to 1 ml PBS prior to being examined using a flow cytometer. Analysis was performed using CellQuest software (Version 5.1, BD Biosciences).
Cd3 antibodies
Manufactured by BD
CD3 antibodies are a type of laboratory equipment used for immunological research and analysis. They are designed to recognize and bind to the CD3 protein complex, which is a key component of the T-cell receptor on the surface of T lymphocytes. CD3 antibodies can be used in various applications, such as cell identification, cell activation, and cell sorting, to study the function and behavior of T cells.
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Lab products found in correlation
2 protocols using cd3 antibodies
1
Quantifying Tumor-Infiltrating CTLs by Flow
2
Suppression Assay of T Cell Proliferation
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Responder cells (CD8+ or CD4+CD25− T cells) were labeled with 2.5 μM CFSE (5 × 106 cell/ml RPMI; 10 min at 37°C under agitation). CFSE labeled cells were then plated onto round bottom 96-well plates coated with CD3 antibodies (8 μg/ml; BD biosciences) in culture medium RPMI- PS-10% heat inactivated FCS, 50 μM β-Mercaptoethanol, 1% non-essential amino acids, and 1% sodium pyruvate. Purified suppressor cells (MDSCs or CD4+CD25+) were added in indicated ratios and plates were incubated at 37°C. The proliferation was measured by assessing dilution of CFSE by flow cytometry after 48 h (CD8+) or 72 h (CD4+CD25−) with CD3/CD28 stimulation (CD28 1 μg/ml). Controls were wells with responder cells without suppressor cells. Treg suppression assay were performed in the presence of IL-2 (50 U/ml).
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