Weakly Stains B220+ Peripheral Blood Leukocytes

March 04, 2016

Immunology
Duke University
Predoctoral

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

eBioscience

Product Name:

Anti-Human/Mouse CD45R (B220) PE

Catalog Number:

12-0452-83

The goal of this experiment was to clearly distinguish B cells in the peripheral blood of female C57Bl/6 mice for subsequent phenotyping. I selected this product because B220 is a B cell-specific surface marker.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Peripheral blood leukocytes

Primary Incubation

Stained cells in 100 uL cold PBS containing Live/Dead and TruStain Fc block (2 ug/mL) on ice for 15 minutes.

Blocking Agent

TruStain Fc block (2 ug/mL)

Secondary Incubation

Added 100 uL cold PBS containing isotype control (4 ug/mL) or CD5 FITC (4 ug/mL) and incubated on ice for 15 minutes.

Tertiary Incubation

NA

Detection

FACS Canto

Results Summary

B220+ B cells peripheral blood leukocytes were distinguishable but not entirely distinct from B220- peripheral blood leukocytes.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

It is relatively cheap as far as antibodies go.

The Bad

It did not complete distinguish B220+ B cells.

The Bottom Line

If you are just generally phenotyping peripheral blood leukocytes, then you can get by with this antibody. However, if you need to very specifically identify B cells for subsequent phenotyping analysis, I would choose a CD19 antibody.

Share your experience with other scientists. Write a Review! »

Join the discussion