Dic n2 plan apo

Manufactured by Nikon

The DIC N2 Plan Apo is a specialized laboratory microscope objective from Nikon. It is designed to provide high-quality, aberration-corrected imaging for a variety of applications.

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Lab products found in correlation

Coolsnap photometrics camera, by Nikon (2 mentions) Eclipse 90i, by Nikon (2 mentions) Cy3 anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions) Ds qi1, by Nikon (1 mentions) Glycerol bp229 1, by Thermo Fisher Scientific (1 mentions) Eclipse 90i microscope, by Nikon (1 mentions) Ds fi3, by Nikon (1 mentions) Eclipse ts2, by Nikon (1 mentions)

4 protocols using dic n2 plan apo

BrdU and Immunofluorescence Staining Protocol

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For BrdU, cells on coverslips were incubated with 1μM BrdU for 30min. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, and then postfixed with 1% PF + 0.01% Tween-20. Coverslips were DNaseI treated for 10min, and the DNaseI reaction was stopped using 20mM EDTA. Coverslips were then blocked with 3% BSA/PBS and incubated in anti-BrdU primary antibody (1:500) followed by incubation in FITC anti-Rat secondary antibody (1:1000). Finally, coverslips were incubated with 0.15 μg/ml DAPI, mounted, and sealed.
For immunofluorescence, cells on coverslips were fixed, permeabilized, and blocked as above and then incubated with the corresponding primary antibodies followed by incubation in FITC anti-rabbit (1:2000) or Cy3 anti-mouse (1:5000) secondary. Finally, coverslips were counterstained with DAPI, mounted, and sealed. All images were acquired at room temperature using a Nikon Eclipse 90i microscope with a 20x/0.17 objective (Nikon DIC N2 Plan Apo) equipped with a CoolSNAP Photometrics camera.

Dahl E.S., Buj R., Leon K.E., Newell J.M., Imamura Y., Bitler B.G., Snyder N.W, & Aird K.M. (2019). Targeting IDH1 as a pro-senescent therapy in high-grade serous ovarian cancer. Molecular cancer research : MCR, 17(8), 1710-1720.

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Immunofluorescence Assay for PML Protein

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Cells (5 × 10⁶/well in 12-well plates) were seeded in 1 mL RPMI-1640 supplemented with 5% FBS overnight and washed with 1x PBS twice on next day. Cells were cultured with 0.5 mL serum-free RPMI for another 24 h Ovcar3 cells were then treated 1 μM KU60019 and 10 μM fenofibrate; Ovcar10 cells were treated 10 μM KU60019 and 25 μM fenofibrate in RPMI + 0.1% FBS for 3 days. KU60019 was administered 5 h prior to fenofibrate. Cells were fixed in 4% paraformaldehyde for 10min and permeabilized in 0.2% Triton X-100 for 5 min. Cells then were stained with PML (1:200, Santa Cruz, Cat# sc-966) in 3% BSA/PBS at room temperature for 1 h. Cells were further incubated with 0.15 μg/ml DAPI in PBS (1 min), mounted, and sealed. Cells were washed three times and then incubated in Cy3 anti-mouse secondary antibody (1:5000, Jackson ImmunoResearch Labs, Cat# 715-165-150) in 3% BSA/PBS at room temperature for 1 h. Finally, cells were incubated with 0.15 μg/ml DAPI in PBS for 1 min, washed three times with PBS, mounted, and sealed. Images were acquired at room temperature using a Nikon Eclipse 90i microscope with a 20x/0.17 objective (Nikon DIC N2 Plan Apo) equipped with a CoolSNAP Photometrics camera.

Chen C.W., Buj R., Dahl E.S., Leon K.E, & Aird K.M. (2020). ATM inhibition synergizes with fenofibrate in high grade serous ovarian cancer cells. Heliyon, 6(9), e05097.

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Immunofluorescence Staining and Imaging Protocol

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Cells were seeded at an equal density on coverslips and fixed with 4% paraformaldehyde. Cells were washed four times with 1× PBS and permeabilized with 0.2% Triton X-100 in PBS for 5 min and then postfixed with 1% paraformaldehyde and 0.01% Tween 20 for 30 min. Cells were blocked for 5 min with 3% BSA/PBS followed by incubation of corresponding primary antibody in 3% BSA/PBS for 1 h at room temperature. Prior to incubation with secondary antibody in 3% BSA/PBS for 1 h at room temperature, cells were washed three times with 1% Triton X-100 in PBS. Cells were then incubated with 0.15 µg/ml DAPI in 1× PBS for 1 min, washed three times with 1× PBS, mounted with fluorescence mounting medium (9 ml of glycerol [BP229-1; Fisher Scientific], 1 ml of 1× PBS, and 10 mg of p-phenylenediamine [PX0730; EMD Chemicals]; pH was adjusted to 8.0–9.0 using carbonate-bicarbonate buffer [0.2 M anhydrous sodium carbonate, 0.2 M sodium bicarbonate]) and sealed. At least 200 cells per coverslip were counted. Images were obtained at room temperature using a Nikon ECLIPSE 90i microscope with a 20×/0.17 objective (Nikon DIC N2 Plan Apo) equipped with a camera (DS-Qi1). Images were acquired using NIS-Elements AR software and processed using ImageJ.

Leon K.E., Buj R., Lesko E., Dahl E.S., Chen C.W., Tangudu N.K., Imamura-Kawasawa Y., Kossenkov A.V., Hobbs R.P, & Aird K.M. (2021). DOT1L modulates the senescence-associated secretory phenotype through epigenetic regulation of IL1A. The Journal of Cell Biology, 220(8), e202008101.

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Senescence-Associated β-Galactosidase Staining

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Cells (5x10 Images were acquired at room temperature using a Nikon Eclipse 90i microscope with a 20x/0.17 objective (Nikon DIC N2 Plan Apo) equipped with a CoolSNAP Photometrics camera.
Senescence-associated beta-galactosidase staining SA-β-Gal staining was performed as previously described (37) . Cells were fixed in 2% formaldehyde/0.2% glutaraldehyde in PBS for 5 min and stained (40 mM Na2HPO4, 150 mM NaCl, 2 mM MgCl2, 5 mM K3Fe(CN)6, 5 mM K4Fe(CN)6, and 1 mg/ml X-gal) overnight at 37 °C in a non-CO2 incubator. Images were acquired at room temperature using an inverted microscope (Nikon Eclipse Ts2) with a 20X/0.40 objective (Nikon LWD) equipped with a camera (Nikon DS-Fi3). Each sample was assessed in triplicate and at least 100 cells per well were counted (> 300 cells per experiment).

Chen C., Buj R., Dahl E.S., Leon K.E., & Aird K.M. (2020). ATM Inhibition Synergizes with Fenofibrate in High Grade Serous Ovarian Cancer Cells.

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