OK Ly6G Antibody, but Has Lot-to-Lot Variability

Department of Medicine
University of Vermont
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Bio X Cell

Product Name:

InVivoMAb anti-mouse Ly6G

Catalog Number:

BE0075

Clone Number:

1A8

I've used this antibody on two different occasions (two years apart), both for neutrophil depletion in C57BL/6 mice. We administered 200 ug in 200 ul of saline 24 hours before, and one hour before lung injury for a total of 400 ug. The first set of experiments worked very well, bringing neutrophils in the BALF from 4x10^6 down to around 0.2x10^6. We went to repeat this two years later and now find ourselves having to retitrate the antibody. Despite all other conditions the same. This is a mild inconvenience, but for the price of the antibody, we should not have to retitrate with every new lot. However, after working with one of the scientists at Bio X Cell, we received a new lot of Ly6G antibody. It is good to know they are responsive to troubleshooting and stand by their product

Experimental Design and Results Summary

Applications

Block/Neutralize/Inhibit

Sample

Mouse C57BL/6

Primary Incubation

200 ug 24h and 1h prior to lung injury

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

N/A

Results Summary

Our first experiments went well using 200ug for 2 injections prior to lung injury. We saw reductions from 4x10^6 to about 0.2x10^6 neutrophils in the BALF. The second attempt at these experiments (two years later) we saw non-significant reductions in BALF neutrophils. 2x10^6 for isotype control, and 1x10^6 for Ly6G.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Works - kind of

The Bad

Expensive with lot-to-lot variability

The Bottom Line

If you have the money, it is a good way to deplete PMNs. You may need to retitrate from lot to lot.

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