CY5-anti-CD3, FITC-anti-CD8, CY5-anti-CD8, CY5-anti-CD4, PE-anti-CD158b (KIR2DS2), PE-anti-NKB1 (KIR3DL1) and FITC-anti-CD28 were obtained from BD PharMingen, San Diego, CA. PE-anti-CD158d (KIR2DL4) was obtained from R&D Systems (Minneapolis, MN) and PE-anti-CD158a/h (KIR2DL1/2DS2) and PE-anti-CD158b1/b2 (KIR 2DL2/2DL3/2DS2) were obtained from Beckman Coulter Immunotech (Fullerton, CA). Five μl of each PE-conjugated anti-KIR Ab were mixed to form a “cocktail” and used to stain the cells (19 (link)). All labeling procedures were performed on ice in PBS containing 10% horse serum and normal human AB serum (Invitrogen) and sodium azide. Cell staining was performed as previously described (20 (link)), and the stained cells were analyzed using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ).
Normal human ab serum
Manufactured by Thermo Fisher Scientific
Normal human AB serum is a biological fluid derived from the blood of individuals with AB blood type. It contains a complex mixture of proteins, nutrients, and other components found in the human circulatory system. This product is commonly used in cell culture and other in vitro applications as a supplement to provide a natural growth environment for cells.
Automatically generated - may contain errors
2 protocols using normal human ab serum
1
Multicolor Flow Cytometry Analysis
2
Multicolor Flow Cytometry Immunophenotyping
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Cy5-conjugated anti-CD3, fluorescein isothiocyanate (FITC)–conjugated anti-CD8, Cy5-conjugated anti-CD8, Cy5-conjugated anti-CD4, phycoerythrin (PE)–conjugated anti-CD158b (killer cell immunoglobulin-like receptor 2DS2 [KIR-2DS2]), PE-conjugated anti-NKB1 (KIR-3DL1), and FITC-conjugated anti-CD28 were obtained from BD PharMingen. PE-conjugated anti-CD158d (KIR-2DL4) was obtained from R&D Systems and PE-conjugated anti-CD158a/h (KIR-2DL1/2DS2) and PE-conjugated anti-CD158b1/b2 (KIR-2DL2/2DL3/2DS2) were obtained from Beckman Coulter Immunotech. Five microliters of each PE-conjugated anti-KIR antibody was mixed to form a cocktail and used to stain the cells (19 (link)). All labeling procedures were performed on ice in PBS containing 10% horse serum and normal human AB serum (Invitrogen) and sodium azide. Cell staining was performed as previously described (20 (link)), and the stained cells were analyzed using a FACSCalibur flow cytometer (BD Biosciences).
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