NK cells and PMN-MDSCs were isolated from PBMCs of mobilized and non-mobilized donors using NK isolation kit and CD66b+ microbeads (purity >98%, data not shown) following manufacturer’s instruction (Miltenyi Biotec, Bergisch Gladbach, Germany). Before starting any experiment, we determined the purity of isolated cells by flow cytometry using anti-CD3-APC, anti-CD19-ECD, and CD56-PC7 (Beckman Coulter, Brea, CA) for NK cells. PMN-MDSCs were labeled with anti-CD3-AF700, anti-CD19-AF700, anti-CD11b-FITC, anti-CD33-PC7, anti-HLA-DR-PE, anti-CD14-ECD, anti-CD45-KrOr, and anti-CD66b-APC desiccated in the Duraclone custom design platform (Beckman Coulter, Brea, CA) adding anti-CD56-BV650 (BioLegend, San Diego, CA) and following manufacturer’s instruction. After the staining procedures, cells were acquired at Cytoflex LX and analyzed with Cytexpert software (v2.4, Beckman Coulter, Brea, CA). Freshly isolated NK cells were immediately used for functional studies and gene expression evaluation.
Anti cd33 pc7
Manufactured by Beckman Coulter
Sourced in Canada
The Anti-CD33-PC7 is a fluorochrome-conjugated monoclonal antibody that binds to the CD33 antigen. CD33 is a cell surface antigen expressed on myeloid progenitor cells and mature myeloid cells. The Anti-CD33-PC7 antibody is used for the identification and enumeration of CD33-positive cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd11b fitc, by Beckman Coulter (2 mentions)
Anti hla dr pe, by Beckman Coulter (1 mentions)
Cd66b microbeads, by Miltenyi Biotec (1 mentions)
Nk isolation kit, by Miltenyi Biotec (1 mentions)
Anti cd3 apc, by Beckman Coulter (1 mentions)
Anti cd56 bv650, by BioLegend (1 mentions)
Anti cd19 ecd, by Beckman Coulter (1 mentions)
Anti cd14 ecd, by Beckman Coulter (1 mentions)
Cytoflex lx, by Beckman Coulter (1 mentions)
Cd56 pc7, by Beckman Coulter (1 mentions)
Cytexpert software, by Beckman Coulter (1 mentions)
Anti cd3 apc a750, by Beckman Coulter (1 mentions)
Anti hla dr pc7, by Beckman Coulter (1 mentions)
Duraclone, by Beckman Coulter (1 mentions)
Anti cd4 apc, by Beckman Coulter (1 mentions)
Anti cd14 pc5, by Beckman Coulter (1 mentions)
Kaluza software, by Beckman Coulter (1 mentions)
Navios flow cytometer, by Beckman Coulter (1 mentions)
2 protocols using anti cd33 pc7
1
Isolation and Characterization of NK Cells and PMN-MDSCs
2
MDSC and T-cell Phenotyping using Flow Cytometry
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Evaluation of MDSC percentage was performed using dry preformulated antibody panels DuraClone (anti-CD11b FITC, anti-CD33 PC7, anti-CD14 PC5.5, anti-human leucocyte antigen locus DR (HLA-DR) Phycoerythrin-Texas Red conjugate, energy coupled dye, cocktail of antibodies anti-CD3 APC-A750, -CD56 APC A750, -CD19 APC A750 (Lin), anti-CD15 pacific blue, and CD45 krome orange, DRAQ-7) from Beckman Coulter. See Figure S1, Supplemental Digital Content 2, https://links.lww.com/QAI/C170 for gating strategy used to identify MDSC.
T-cell phenotype was accomplished using dry preformulated antibody panels DuraClone (anti-CD3 APC-A750, anti-CD4 APC, anti-CD197 PE, anti-CD45RA Phycoerythrin-Texas Red conjugate, energy coupled dye, anti-HLA-DR PC7, anti-CD38 pacific blue, anti-CD45 krome orange) from Beckman Coulter. Single staining and compensation controls were used to set up flow cytometry experiments. Acquisition of 100,000 events was performed in the leukocyte-gated population on Navios flow cytometer and analyzed with Kaluza software (Beckman Coulter).
T-cell phenotype was accomplished using dry preformulated antibody panels DuraClone (anti-CD3 APC-A750, anti-CD4 APC, anti-CD197 PE, anti-CD45RA Phycoerythrin-Texas Red conjugate, energy coupled dye, anti-HLA-DR PC7, anti-CD38 pacific blue, anti-CD45 krome orange) from Beckman Coulter. Single staining and compensation controls were used to set up flow cytometry experiments. Acquisition of 100,000 events was performed in the leukocyte-gated population on Navios flow cytometer and analyzed with Kaluza software (Beckman Coulter).
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