Lonza ACK Lysis Buffer in the lysis of red blood cells

March 24, 2015

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Company:

Lonza

Product Name:

ACK Lysis Buffer

Catalog Number:

10-548e

My work focuses on the effects of immunomodulation in Parkinson's disease. Here, Lonza's ACK Lysis Buffer is used to lyse red blood cells from splenocytes.

Experimental Design and Results Summary

Application

Cell culture

Starting Material

Splenocytes

Protocol Overview

Spleens are removed from mice, made into a single cell suspension, and 1ml of ACK lysis buffer is added for every spleen removed. Mixture is then left in a 37 degree waterbath for 2 minutes with gentle mixing every minute. Q.S. solution up to 50ml and centrifuge to remove the ACK lysis buffer from cells.

Tips

Sometimes the cells will need to incubate for longer than 2 minutes with the ACK Lysis Buffer

Results Summary

Visually you can see after you centrifuge that the red blood cells are gone, and this can be verified through trypan blue staining.

Additional Notes

If red blood cells still present after incubation and centrifugation, another round of incubation with the ACK lysis buffer can be preformed, but it may yeild a lower cell count in the end.

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Summary

The Good

Very easy way to remove red blood cells from your culture.

The Bad

All the red blood cells arent always lysed with the first round of incubation in ACK lysis buffer

The Bottom Line

The Lonza ACK lysis buffer provides an easy way to remove red blood cells from culture, but has the potential to lower cell yield if not used correctly.

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