Mouse anti tra 1 60

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Mouse anti-Tra-1–60 is a monoclonal antibody that specifically binds to the Tra-1-60 antigen, a cell surface marker expressed on embryonic stem cells and induced pluripotent stem cells. It can be used for the detection and characterization of these cell types.

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Close spelling variants of this product

Mouse anti-TRA-1-60-PE Anti-TRA-1-60 TRA-1-60

4 protocols using mouse anti tra 1 60

Immunocytochemical Characterization of Cells

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Cells were grown on precoated glass coverslips (Neuvitro Corporation, Vancouver, WA) and fixed in 4% paraformaldehyde solution in phosphate-buffered saline. Cells were blocked with bovine serum albumin and normal serum from the species specific to the primary antibodies. Primary antibodies used were mouse IgG2b anti-TuJ1 (Sigma), chicken anti–glial fibrillary acidic protein (GFAP; Sigma), rabbit anti-S100ß (Dako Corporation, Carpinteria, CA), guinea pig anti-vGluT1 (Synaptic Systems, Göttingen, Germany), mouse anti–Tra-1–60 (Thermo Fisher Scientific), goat anti-Oct3/4 (Santa Cruz Biotechnology, Santa Cruz, CA), and rabbit anti-Sox2 (Millipore). Fluorophore-conjugated donkey secondary antibodies against the primary antibody host species IgG were from Thermo Fisher Scientific. Nuclei were counterstained with DAPI (Thermo Fisher Scientific), and cells were mounted using Prolong Gold media (Thermo Fisher Scientific). Cell counts were performed in ImageJ, and values for each genotype and patient background were determined by the average of 3 fields per coverslip and 3 coverslips.

Wadhwani A.R., Affaneh A., Van Gulden S, & Kessler J.A. (2019). Neuronal Apolipoprotein E4 Increases Cell Death and Phosphorylated Tau Release in Alzheimer Disease. Annals of neurology, 85(5), 726-739.

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Immunofluorescence Staining Protocol

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The following antibodies were used: rabbit anti-GFAP (DAKO Agilent, Santa Clara, CA, Z0334), rabbit anti-caspase-6 (Cell Signaling Technologies, Danvers, MA, 9762), rabbit anti-Caspase-6 (abcam, Cambridge, UK, ab185645), rabbit anti-D225 (Chen et al., 2013 (link)), mouse anti-GFAP (Sigma, GA5), mouse anti-pSer13-GFAP (KT13 [Sekimata et al., 1996 (link)]), mouse anti-pan Actin, mouse anti-Tra-1–60, mouse anti-SSEA4, rabbit anti-Oct4, rabbit anti-Sox2, and Alexa 488- and Alexa 594-congujated goat anti mouse or rabbit antibodies (Thermo Fisher Scientific, Waltham, MA).

Battaglia R.A., Beltran A.S., Delic S., Dumitru R., Robinson J.A., Kabiraj P., Herring L.E., Madden V.J., Ravinder N., Willems E., Newman R.A., Quinlan R.A., Goldman J.E., Perng M.D., Inagaki M, & Snider N.T. (2019). Site-specific phosphorylation and caspase cleavage of GFAP are new markers of Alexander disease severity. eLife, 8, e47789.

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Comprehensive Immunofluorescence and Western Blot Antibody Protocol

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The following primary antibodies and concentrations were used in this study: rabbit anti-GFAP (DAKO, Agilent, clone Z0334; IF 1:500, WB 1:10,000), mouse anti-GFAP (Sigma, clone GA5; IF 1:300), mouse anti-Gigaxonin (Santa Cruz Biotechnology, F3, WB 1:200), rabbit anti-Vimentin (Cell-Signaling Technology, D21H3, IF 1:100), mouse anti-Vimentin (Thermo Fisher Scientific, V9, WB 1:1000), mouse anti-Keratin 8 (Thermo Fisher Scientific, TS1, IF 1:100), rat anti-K8 (Developmental Studies Hybridoma Bank, Troma I, WB 1:5000), rabbit anti-Lamin A/C (Santa Cruz Biotechnology), rabbit anti-Lamin B1 (Abcam ab16048, IF 1:10,000, WB 1:10,000), mouse anti-Nestin (Thermo Fisher Scientific, 10C2, IF 1:200), mouse anti-NF-M/H (clone RMdO-20; IF 1:100), mouse anti-Tra-1-60 (Thermo Fisher Scientific, 41-1000, IF 1:300), mouse anti-Tra-1-81 (Thermo Fisher Scientific, 41-1100, IF 1:300), rabbit anti-Oct4 (Abcam, ab19857, IF 1:40), and rabbit anti-Sox2 (Thermo Fisher Scientific, 48-1400, IF 1:125). The following secondary antibodies and concentrations were used: Alexa 488- and Alexa 594-conjugated goat-, anti-mouse-, and rabbit-antibodies (Thermo Fisher Scientific, IF 1:500), and peroxidase-conjugated goat-, anti-mouse-, and rabbit-antibodies (Sigma, WB 1:5000).

Battaglia R.A., Faridounnia M., Beltran A., Robinson J., Kinghorn K., Ezzell J.A., Bharucha-Goebel D., Bönnemann C.G., Hooper J.E., Opal P., Bouldin T.W., Armao D, & Snider N.T. (2023). Intermediate filament dysregulation in astrocytes in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN). Molecular Biology of the Cell, 34(12), ar121.

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Comprehensive Antibody Panel for Cellular Characterization

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The following primary antibodies and concentrations were used in this study: rabbit anti-GFAP (DAKO, Agilent, clone Z0334; IF 1:500, WB 1:10,000), mouse anti-GFAP (Sigma, clone GA5; IF 1:300), mouse anti-pSer13-GFAP (gift from Dr. Masaki Inagaki, clone KT13, IF 1:20), mouse anti-Gigaxonin (Santa Cruz Biotechnology, F3, WB 1:200), rabbit anti-Vimentin (Cell Signaling Technology, D21H3, IF 1:100), mouse anti-Vimentin (Thermo Fisher Scientific, V9, WB 1:1000), mouse anti-Keratin 8 (Thermo Fisher Scientific, TS1, IF 1:100), rat anti-K8 (Developmental Studies Hybridoma Bank, Troma I, WB 1:5000), rabbit anti-Lamin A/C (Santa Cruz Biotechnology), rabbit anti-Lamin B1 (Abcam ab16048, IF 1:10,000, WB 1:10,000), mouse anti-Nestin (Thermo Fisher Scientific, 10C2, IF 1:200), mouse anti-Tra-1–60 (Thermo Fisher Scientific, 41–1000, IF 1:300), mouse anti-Tra-1–81 (Thermo Fisher Scientific, 41–1100, IF 1:300), rabbit anti-Oct4 (Abcam, ab19857, IF 1:40), and rabbit anti-Sox2 (Thermo Fisher Scientific, 48–1400, IF 1:125). The following secondary antibodies and concentrations were used: Alexa 488- and Alexa 594-conjugated goat anti mouse and rabbit antibodies (Thermo Fisher Scientific, IF 1:500), and peroxidase-conjugated goat anti-mouse and rabbit antibodies (Sigma, WB 1:5000).

Battaglia R., Faridounnia M., Beltran A., Robinson J., Kinghorn K., Ezzell J.A., Bharucha-Goebel D., Bonnemann C., Hooper J.E., Opal P., Bouldin T.W., Armao D, & Snider N. (2023). Intermediate filament dysregulation and astrocytopathy in the human disease model of KLHL16 mutation in giant axonal neuropathy (GAN). bioRxiv.

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