IgM-PB Works Great On Human PBMC

Medicine
Vanderbilt University Medical Center
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Pacific Blue™ anti-human IgM Antibody

Catalog Number:

314514

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 IgM-PB was because it fit well in my 14-parameter panel with minimal compensation issues with other antibody conjugates.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Cryopreserved human PBMC

Primary Incubation

0.1uL (1:2000) IgM-PB in 200uL FACS buffer with 2x10^6 cells; 15 minutes; room temperature

Blocking Agent

None

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD LSRII 5-laser Flow Cytometer

Results Summary

A single color titration of IgM-PB was performed and 0.1uL (1:2000) yielded the highest stain index. We routinely use IgM-PB 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 surface antibodies, including IgM-PB. ~2 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. It detects the expected frequency of IgM+ B cells in human PBMCs.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Staining pattern is good, really economical (because you only use at 1:2000!)

The Bad

None

The Bottom Line

Antibody works great!

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