One of the few LAP antibodies that work

March 05, 2015

Neurology
Brigham and Women's Hospital
Postdoc

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Company:

BioLegend

Product Name:

Brilliant Violet 421™ anti-mouse LAP (TGF-β1) Antibody (clone: TW7-16B4)

Catalog Number:

141407

Info: View Product View Product Specs

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Our group is interested in regulatory Th3 cells that express membrane bound TGF-beta. The only way to detect these cells is by surface staining of the TGF-beta complex by antibodies recognizing latency associated peptide (LAP) that keeps TGF-beta in an inactive state. We compared different antibodies and found that the best way to detect membrane bound TGF-beta is the Brilliant Violet 421™ anti-mouse LAP (TGF-β1) Antibody.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse spleen cells

Primary Incubation

1/150, 50ul, 30 min, 4 degrees

Blocking Agent

Fc-block (anti mouse CD16/CD32, 1/200, 15 min before putting the LAP antibody)

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometry (LSRII)

Results Summary

Normally 3-8% of splenic CD4+ T cells are positive for LAP / membrane TGF-beta. An example for a normal staining is shown in the figure. We always use an FMO control (all antibodies minus the LAP antibody) to better adjust the gates).

Additional Notes

We only have good results with the LAP antibody conjugated to BV-421 or PE. Cells can be fixed after staining but permeabilization is not recommanded as it deteriorates the staining.

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Summary

The Good

Bright staining, the best LAP antibody on the market as far as our group knows.

The Bad

Only BV-421 and PE conjugated antibodies give satisfactory results

The Bottom Line

The best LAP antibody on the market as far as our group knows, but only BV-421 and PE conjugated antibodies give satisfactory results. Cells can be fixed after staining but permeabilization is not recommanded as it deteriorates the signal.

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