Anti pdpn clone nz 1

Manufactured by Thermo Fisher Scientific

Anti-Pdpn: clone NZ-1.3 is a monoclonal antibody that recognizes the podoplanin (Pdpn) protein. Pdpn is a transmembrane glycoprotein expressed on the surface of various cell types, including lymphatic endothelial cells, type I alveolar cells, and certain tumor cells. The NZ-1.3 clone is a research-use antibody that can be used to detect and study the expression of Pdpn in different biological samples and experimental settings.

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Lab products found in correlation

Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Anti cd4 clone a161a1, by BioLegend (1 mentions) Anti cd28 37, by BioLegend (1 mentions) Anti cd8 rm2206, by BioLegend (1 mentions) Anti cd3ε 145 2c11, by BD (1 mentions) Anti thy1 clone 5e10, by BD (1 mentions) Anti human cd3 clone okt3, by Thermo Fisher Scientific (1 mentions) Anti human cd28 clone cd28, by Thermo Fisher Scientific (1 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)

3 protocols using anti pdpn clone nz 1

Comprehensive Murine Tissue Analysis Panel

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For flow cytometry and imaging studies with murine tissues, the following antibodies were purchased from Biolegend: anti-CD45 (30-F11), anti-CD31 (390), anti-PDPN (8.1.1), anti-α-smooth muscle actin (1A4), anti-Thy1 (53–2.1), anti-CD140α (APA5), anti-CD140b (APB5), anti-CD106 (429), anti-CD4 (RM4.5), and anti-CD8 (RM2206). The anti-FAP used for flow cytometry analysis was provided by Ellen Pure and has been validated (anti-mouse FAP 73.3)(8 (link)). For immunofluorescent analysis, the Roche anti-FAP 28H1 clone was used (26 (link)). Functional-grade purified anti-CD3ε (145–2C11) was from BD Biosciences, and anti-CD28 (37.51) was from Biolegend. Antibodies used to stain human samples include viability dye (L10119; Life Technologies), anti-PDPN (clone NZ1.3; eBioscience), anti-Thy1 (clone 5E10; BD Biosciences), anti-CD4 (clone A161A1; Biolegend) and anti-CD8 (clone HIT8α; Biolegend). For PBMC stimulation, anti-human CD3 (clone OKT3; eBioscience) and anti-human CD28 (clone CD28.2; eBioscience) were used. L-NMMA (NG-monomethyl-L-arginine) was purchased from Calbiochem and CFSE (carboxyfluorescein diacetate succinimidyl ester) was from Invitrogen.

Cremasco V., Astarita J., Grauel A.L., Keerthivasan S., MacIsaac K., Woodruff M.C., Wu M., Spel L., Santoro S., Amoozgar Z., Laszewski T., Migoni S.C., Knoblich K., Fletcher A.L., LaFleur M., Wucherpfennig K.W., Pure E., Dranoff G., Carroll M.C, & Turley S.J. (2018). FAP delineates heterogeneous and functionally divergent stromal cells in immune-excluded breast tumors. Cancer immunology research, 6(12), 1472-1485.

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Single-cell tissue immunophenotyping

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Single-cell suspensions obtained from tissue digests were blocked in PBS 5% BSA containing human anti-Fc block (Miltenyi), except when CD16 was in the panel, stained for FACS analysis or sorting with Fixable Viability Dye eFluor 780 (eBioscience, 1:1,000), DAPI (Invitrogen, 1:50,000) and antibodies (all from Biolegend, except anti-Pdpn: clone NZ-1.3 from eBioscience and anti-FAP:sheep from Biotechne) in PBS with 5% BSA and 5 mM EDTA for 20 min on ice. After washing in the same buffer, cells were either analyzed (LSRII or Fortessa X20) or sorted (Aria III, 100-µm nozzle). All antibodies used for FACS analysis and sorting can be found in Supplementary Table 10.

Friedrich M., Pohin M., Jackson M.A., Korsunsky I., Bullers S.J., Rue-Albrecht K., Christoforidou Z., Sathananthan D., Thomas T., Ravindran R., Tandon R., Peres R.S., Sharpe H., Wei K., Watts G.F., Mann E.H., Geremia A., Attar M., McCuaig S., Thomas L., Collantes E., Uhlig H.H., Sansom S.N., Easton A., Raychaudhuri S., Travis S.P, & Powrie F.M. (2021). IL-1-driven stromal–neutrophil interactions define a subset of patients with inflammatory bowel disease that does not respond to therapies. Nature Medicine, 27(11), 1970-1981.

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Single-cell FACS Staining and Sorting

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Single-cell suspensions obtained from tissue digests were stained for FACS analysis or sorting with antibodies (all from Biolegend, except anti-Pdpn: clone NZ-1.3 from eBioscience) in PBS with 5% BSA and 5mM EDTA for 20 minutes on ice. After washing in the same buffer, cells were analysed (LSRII or Fortessa X20) or sorted (Aria III, 100um nozzle).

Friedrich M., Pohin M., Jackson M., Korsunsky I., Bullers S.J., Rue-Albrecht K., Christoforidou Z., Sathananthan D., Ravindran R., Peres R., Sharpe H., Wei K., Watts G.F., Mann E.H., Geremia A., Thomas T., Attar M., McCuaig S., Thomas L., Collantes E., Uhlig H.H., Sansom S.N., Easton A., Raychaudhuri S., Travis S., & Powrie F. (2021). IL-1-driven stromal-neutrophil interaction in deep ulcers defines a pathotype of therapy non-responsive inflammatory bowel disease.

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