1A8 Depletion of PMN

April 26, 2016

Department of Pediatrics
University of California, San Diego
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioXCell

Product Name:

In Vivo MAb anti-mouse Ly6G

Catalog Number:

BE0075-1

This antibody was used to deplete PMN from mice via interperitoneal injeciton.

Experimental Design and Results Summary

Applications

Block/Neutralize/Inhibit

Sample

Mice (analysis of depletion done in Mtb infected lungs)

Primary Incubation

NA

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Loss of Gr-1 high cells

Results Summary

Inject mice intraperitoneally with 200ng 1A8 every 48 hrs - done from 10 - 28 days post infection with M. tuberculosis. Analyzed lungs at 21 days post infection for loss of Gr-1 high cells. Left shows a control (IgG) treated mouse, the right shows a 1A8 treated mouse. I also showed flow analysis of Ly6G levels - this is misleading because of epitope blocking, but the Gr-1 stainings shows loss of PMN.

Additional Notes

None

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Summary

The Good

This antibody works well, there are no other options in terms of PMN depletion Ab clones that I am aware of.

The Bad

Expensive - can be produced from the hybridoma, if you get an MTA from Dr. Malek but the cells can be difficult to culture.

The Bottom Line

Best option for in vivo PMN depletion. Must treat every 48 hours, and check depletion at different time points, the Gr-1 Ab will develop resistance in vivo within a few days, the Ly6G (1A8) Ab seems to work longer (at least 2 weeks of treatment) - but need to check in specific organ and timepoint to verify.

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