Miltenyi's Anti-TNFRH3 for Flow Cytometry

October 18, 2019

Biological Sciences Platform
Sunnybrook Research Institute
Postdoctoral Researcher

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Company:

Miltenyi Biotec

Product Name:

Anti-TNFRH3 antibody FITC

Catalog Number:

130-115-897

Clone Number:

REA953

To validate this antibody in flow cytometry it was used to stain HEK293T cells transfected with a plasmid encoding mTNFRSF26 from Origene. As a negative control, non-transfected HEK293T cells were also used. The antibody was used at a dilution of 1:50. The isotype FITC antibody used was isotype control REA293 (cat no 130-113-449).The REA953 stained the transfected HEK293T cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

HEK293T transfectants

Primary Incubation

1:50 dilution 1 hour incubation on ice

Blocking Agent

BSA

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow cytometry

Results Summary

The antibody positively stained the transfected HEK293T cells.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Cheap and a range of 3 fluors available

The Bad

As it is a recombinant human antibody, you have to buy the isotype control from MACS. May non-specifically bind to a human protein, as human cells do not have a TNFRSF26 gene but there is binding on non transfected HEK293T cells.

The Bottom Line

A potentially good antibody for flow cytometry staining of TNFRSF26. Will need to be further validated in mouse cells, ideally using a KO mouse line or KO mouse.

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