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Cd14 and cd15 positive selection kits cd14 cd15

Manufactured by Miltenyi Biotec

The CD14 and CD15 positive selection kits are laboratory tools used to isolate specific cell populations from biological samples. The kits utilize antibodies coupled to magnetic beads to selectively bind and separate CD14-positive and CD15-positive cells, enabling the user to isolate these cell types for further analysis or experimentation.

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2 protocols using cd14 and cd15 positive selection kits cd14 cd15

1

Purification and Functional Evaluation of gMDSC

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gMDSC were enriched or depleted using sequential magentic bead isolation: CD14 and CD15 positive selection kits (CD14CD15+) (Miltenyi Biotec) as per the manufacturer’s instructions. In selected cases gMDSC were highly purified by flow cytometry on the basis of CD11b, CD14 and CD15 expression on a FACSAria (BD Bioscience). Where indicated PBMC or PBMC enriched/depleted of gMDSC were stimulated with either: 0.5 μg/ml of an HLA-A and HLA-B-restricted peptide pool spanning the immune-dominant proteins of CMV, EBV and influenza (CEF) (JPT Peptide Technologies), 1 μg/ml overlapping peptides (pool of 15-mer peptides overlapping by 10 residues) spanning the core of HBV genotype D (JPT Peptide Technologies), 1 μg/ml HLA-A2-restricted NLVPMVATV peptide from CMV pp65 (ProImmune) or 0.5 μg/ml plate-bound anti-CD3 and 0.5 μg/ml anti-CD28 (eBioscience). All cultures were carried out in the presence of 20 IU/ml recombinant human IL-2 (Miltenyi Biotec) in cRPMI (Life Technologies) for 5-7 days at 37 °C. Specific responses were detected by intracellular cytokine staining after re-stimulation on day 4/6 in the presence of 1 μg/ml brefeldin-A (BFA, Sigma-Aldrich). Where indicated, 0.5 mM N-hydroxyl-nor-L-arginine (nor-NOHA) (Calbiochem), an arginase I specific inhibitor, was added to the culture on day 0.
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2

Purification and Functional Evaluation of gMDSC

Check if the same lab product or an alternative is used in the 5 most similar protocols
gMDSC were enriched or depleted using sequential magentic bead isolation: CD14 and CD15 positive selection kits (CD14CD15+) (Miltenyi Biotec) as per the manufacturer’s instructions. In selected cases gMDSC were highly purified by flow cytometry on the basis of CD11b, CD14 and CD15 expression on a FACSAria (BD Bioscience). Where indicated PBMC or PBMC enriched/depleted of gMDSC were stimulated with either: 0.5 μg/ml of an HLA-A and HLA-B-restricted peptide pool spanning the immune-dominant proteins of CMV, EBV and influenza (CEF) (JPT Peptide Technologies), 1 μg/ml overlapping peptides (pool of 15-mer peptides overlapping by 10 residues) spanning the core of HBV genotype D (JPT Peptide Technologies), 1 μg/ml HLA-A2-restricted NLVPMVATV peptide from CMV pp65 (ProImmune) or 0.5 μg/ml plate-bound anti-CD3 and 0.5 μg/ml anti-CD28 (eBioscience). All cultures were carried out in the presence of 20 IU/ml recombinant human IL-2 (Miltenyi Biotec) in cRPMI (Life Technologies) for 5-7 days at 37 °C. Specific responses were detected by intracellular cytokine staining after re-stimulation on day 4/6 in the presence of 1 μg/ml brefeldin-A (BFA, Sigma-Aldrich). Where indicated, 0.5 mM N-hydroxyl-nor-L-arginine (nor-NOHA) (Calbiochem), an arginase I specific inhibitor, was added to the culture on day 0.
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