Anti-Human CD19-BUV395 Is Great Option For Multi-Parameter Panels

Medicine
Vanderbilt University Medical Center
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

Anti-human CD19-BUV395 (clone SJ25C1)

Catalog Number:

BD 563551

The general aim of our research is to characterize the phenotype and function of pathogenic B cells in various diseases. We are building fluorescence and mass cytometry panels to study human B cells in the context of autoimmune and idiopathic pulmonary diseases. The main reason I selected CD19-BUV395 is because there are little to no compensation issues between BUV395 and the rest of our panel; and since most markers in the panel are B cell markers, this was especially important.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Cryopreserved human PBMC

Primary Incubation

1uL CD19-BUV395 in 200uL FACS buffer with 10^6 cells; 15 minutes; room temperature

Blocking Agent

None.

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

BD LSRII 5-laser Flow Cytometer

Results Summary

A single color titration of CD19-BUV395 was performed and 1uL CD19-BUV395 yielded the highest stain index. We routinely use CD19-BUV395 in our 14-parameter human B cell panel to identify B cells in patient samples with various diseases. Cyropreserved human PBMCs are thawed, washed twice in PBS, and stained with a cocktail of antibodies including CD19-BUV395. 1 million cells in 200uL FACS buffer are incubated with antibodies for 15 minutes at room temperature.As evidenced in the attached figure, the staining looks great and the frequency detected is within the expected range for circulating human B cells.

DOI or PMID #

N/A

Additional Notes

There is slight bleedover between the BUV395 and PacBlue channels. Nothing major, but good to be aware of.

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Summary

The Good

Staining pattern is great, detects expected % of B cells

The Bad

It's expensive.

The Bottom Line

It's expensive, but works great and is well worth the cost when working with a full (14+ parameter) fluorescence cytometry panel.

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