Cells were stained with cell-surface antibodies against CD3-allophycocyanin-Cy7 (clone SK7, BD Pharmingen, San Jose, USA), CD4-phycoerythrin (clone RPA-T4, BD Pharmingen), CD8-fluorescein isothiocyanate (clone RPA-T8; BD Pharmingen) and CD33-allophycocyanin (clone WM53, BD Pharmingen). Cells were washed twice with flow cytometry staining buffer and re-suspended in Pre-Sort buffer (BD Biosciences). 7-AAD viability dye (eBioscience, San Diego, USA) was used to gate live cells. BD FACSAria III SORP cell sorter on BD FACSDiva software (BD Biosciences) was used for sorting pure CD4+ (7AAD–CD3+CD4+CD8–CD33–), CD8+ (7AAD–CD3+CD4–CD8+CD33–) and CD33+(7AAD–CD3–CD4–CD8–CD33+) populations. Applicable measures were taken to ensure minimal sorter-induced cell stress (SICS). Data analyses were performed on FlowJo V10 software (FlowJo, Ashland, USA).
Cd33 allophycocyanin clone wm53
Manufactured by BD
Sourced in United States
CD33-allophycocyanin (clone WM53) is a fluorescent-labeled antibody used in flow cytometry applications. It binds to the CD33 antigen, which is expressed on the surface of myeloid cells.
Automatically generated - may contain errors
Lab products found in correlation
7 aad viability dye, by Thermo Fisher Scientific (2 mentions)
Pre sort buffer, by BD (2 mentions)
Cd3 allophycocyanin cy7 clone sk7, by BD (2 mentions)
Facsaria 3 sorp cell sorter on facsdiva software, by BD (1 mentions)
Cd4 phycoerythrin clone rpa t4, by BD (1 mentions)
Cd8 fluorescein isothiocyanate clone rpa t8, by BD (1 mentions)
Facsaria 3 sorp cell sorter, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Fcr blocking reagent, by Miltenyi Biotec (1 mentions)
2 protocols using cd33 allophycocyanin clone wm53
1
Fluorescence-Activated Cell Sorting of Immune Cell Subsets
2
Single-cell Sorting of CD33+ Monocytes
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Single-cell suspensions from TT were resuspended in 100-µl flow cytometry staining buffer (PBS with 1% FCS and 0.1% sodium azide). Fc receptors (FcR) were blocked using FcR Blocking Reagent (Miltenyi Biotech, Bergisch Gladbach, Germany), and 7-AAD viability dye (eBioscience, San Diego, USA) was used to gate live cells. Cells were stained with cell surface antibodies against CD3-allophycocyanin-Cy7 (clone SK7, BD Pharmingen, San Jose, USA) and CD33-allophycocyanin (clone WM53, BD Pharmingen). Cells were washed twice with flow cytometry staining buffer and re-suspended in Pre-Sort buffer (BD Biosciences). BD FACSAria III SORP cell sorter on BD FACSDiva software (BD Biosciences) was used for sorting pure CD33+(7AAD–CD3–CD33+) populations. Relevant procedures were followed to ensure minimum sorter-induced cell stress (SICS), as previously described [15 (link)]. Flow cytometric representations were performed on FlowJo V10 software (FlowJo, Ashland, USA).
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