Getting Rid Of Red Blood Cells

University of California San Francisco
Medicine
Research Associate

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Company:

Sigma

Product Name:

Red Blood Cell Lysing Buffer Hybri-Max

Catalog Number:

R7757-100ml

This product is great. It gently gets rid of red blood cell contamination in my lung epithelial preps.

Experimental Design and Results Summary

Application

Red Blood Cell Lysing buffer

Starting Material

Mouse Epithelial Cells

Protocol Overview

Mouse lungs were removed and incubated in 15U/ml Dispase for 45 mins rocking at room temp. In a cell culture hood add 10ml of sort buffer and 50U/ml DNAse. Pipet up and down until the lung tissue is dissociated. Incubate 10 mins rocking at 37 degrees. Transfer the contents through 100µm, 70µm, and 40µm cell strainers over 50ml tubes. Transfer filtered suspension to a 15mL tube. Spin for 5 min at 550g at 4°C to pellet cells. Resuspend in 1ml Hybri-Max Red Blood Cell lysing buffer per 100million cells and rock gently for one minute. Top off with PBS and spin for 5 min at 550g at 4°C Resuspend in 10ml of sort buffer and let recover shaking for at least one hour at 37°C.

Tips

I usually filter my cells again after the lysis to get rid of extra debris created by lysing step.

Results Summary

This product nicely gets rid of all red blood cells which is helpful for downstream applications such as flow cytometry and cell culture.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

This product is inexpensive and gentle on other cell types.

The Bad

Nothing!

The Bottom Line

I've tried other methods of removing red blood cells from my cell preps and most of them were very rough on my cells. This product is the best in my opinion.

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