PE Mouse Anti-Human CD138

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	, approximately 100 μL of EDTA‐anticoagulated pleural fluid samples were immunophenotyped using six combinations of monoclonal antibodies and a direct immunofluorescence.	The cells were acquired in a FACSCanto flow cytometer (Becton Dickinson, San Jose, CA) using the FACSDiva software (Becton Dickinson). A minimum of 100,000 events were acquired. For data analysis the Infinicyt software (Cytognos SL, Salamanca, Spain) was used.

Protocol tips
, approximately 100 μL of EDTA‐anticoagulated pleural fluid samples were immunophenotyped using six combinations of monoclonal antibodies and a direct immunofluorescence.
Downstream tips
The cells were acquired in a FACSCanto flow cytometer (Becton Dickinson, San Jose, CA) using the FACSDiva software (Becton Dickinson). A minimum of 100,000 events were acquired. For data analysis the Infinicyt software (Cytognos SL, Salamanca, Spain) was used.

Publication protocol

A six‐color flow cytometric immunophenotyping study of the pleural fluid was carry out. Briefly, approximately 100 μL of EDTA‐anticoagulated pleural fluid samples were immunophenotyped using six combinations of monoclonal antibodies and a direct immunofluorescence stain‐and‐then‐lyse technique—(Fluorescein isothiocyanate (FITC)/phycoerythrin (PE)/peridin chlorophyll protein cyanin 5.5 (PerCP‐Cy5.5)/PE‐cyanin 7 (PE‐Cy7)/allocycocyanin (APC)/APC‐cyanin 7(APC‐Cy7): 1CD7/CD3/CD5/CD8/CD4/CD45; 2 Kappa/Lambda/CD19/‐/CD45/CD20; 3 CD38/CD138/CD19/CD56/CD45/CD20; 4 CD38/CD30/HLA‐DR/CD10/CD45/CD20; 5 CD38/cyCD79a/CD19/CD10/CD45/CD20; 6 CD38/CD138/CD117/CD28/CD45/CD81; and 7 cyKappa/cyLambda/CD19/CD56/CD45/CD38. (Monoclonal antibodies clones and companies are described in Table 1). The cells were acquired in a FACSCanto flow cytometer (Becton Dickinson, San Jose, CA) using the FACSDiva software (Becton Dickinson). A minimum of 100,000 events were acquired. For data analysis the Infinicyt software (Cytognos SL, Salamanca, Spain) was used.

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