BD BrdU Flow Kit Review

August 08, 2013

BlueskyReddit

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review
UCLA
Molecular and Medical Pharmacology
Postdoc
Company: BD Biosciences
Product Name: FITC BrdU Flow Kit
Catalog Number: 559619


Info: View Product View Product Specs

Our lab investigates proliferation and cell cycle in murine hematopoietic cells.  To do this we perform flow cytometry on cells stained with antibodies against cell surface markers and labeled with BrdU and DAPI.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Mouse lymphocytes

Protocol Overview

I labeled mouse lymphocytes with BrdU in vitro for 2h. I collected the cells, stained with surface antibodies, then performed fixation and permeabilization, DNase treatment, and intra-cellular staining for BrdU. My goal was to determine cell cycle status of mouse T lymphocytes in the context of surface marker expression.

Tips

Used DAPI instead of 7AAD for DNA content

Results Summary

BrdU incorporated into cycling cells as demonstrated by population that was positive for BrdU by flow cytometry (50%). No BrdU+ population observed with sample that was unlabeled with BrdU (not incubated with BrdU in vitro, but rest of protocol the same) -- very low background, no non-specific antibody binding. DAPI used on linear scale to show DNA content on X axis with BrdU levels on Y axis and characteristic BrdU horseshoe shape obtained.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Nice signal, low background with antibody. Flexibility in steps of protocol to leave overnight.

The Bad

Kit is expensive and some components run out before others. Individual items are expensive to purchase separately.

The Bottom Line

I would recommend for assessment of cells that are undergoing DNA synthesis/proliferating cells/identifying cells that are actively cycling.

Join the discussion