Immunofluorescence Staining of Spheroids

Spheroids were plated onto glass 22mm circular coverslips and grown until mature. Coverslips were then fixed in cold 2% paraformaldehyde for 15 minutes and rinsed in PBS. H&E stains were performed as previously described (20 (link)). IF was performed as previously described (20 (link)). A conjugated antibody against Ki67 (Cell Signaling #11882 [1:50] (488 conjugate)) was used and slides were mounted with Prolong gold DAPI mounting media (Invitrogen #P36931).

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Fricke S.L., Payne S.N., Favreau P.F., Kratz J.D., Pasch C.A., Foley T.M., Yueh A.E., Van De Hey D.R., Depke M.G., Korkos D.P., Sha G.C., DeStefanis R.A., Clipson L., Burkard M.E., Lemmon K.K., Parsons B.M., Kenny P.A., Matkowskyj K.A., Newton M.A., Skala M.C, & Deming D.A. (2018). MTORC1/2 inhibition as a therapeutic strategy for PIK3CA mutant cancers. Molecular cancer therapeutics, 18(2), 346-355.

Publication 2018

Prolong gold DAPI mounting media

Antibody Cold Dapi Gold Paraformaldehyde Stains

Corresponding Organization : University of Wisconsin Carbone Cancer Center

Other organizations : Morgridge Institute for Research, Gundersen Health System, William S. Middleton Memorial Veterans Hospital

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Variable analysis

independent variables

Spheroids were plated onto glass 22mm circular coverslips

dependent variables

H&E stains
IF (Immunofluorescence) with Ki67 (Cell Signaling #11882 [1:50] (488 conjugate))

control variables

Coverslips were fixed in cold 2% paraformaldehyde for 15 minutes
Coverslips were rinsed in PBS
Slides were mounted with Prolong gold DAPI mounting media (Invitrogen #P36931)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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