Ki-67 immunostaining was conducted on tumors from treprostinil treated and control animals as as previously described(22 (link), 23 (link)). In brief, 5 micron lung sections from the urethane, ET, and LT groups were deparaffinized, peroxidases blocked with 3% hydrogen peroxide, and antigen retrieval performed in boiling Diva Decloaker (Biocare Medical, DV2004G1) reagent under pressure for 10 min. Sections were incubated with Ki67 primary Ab (1:25 dilution, Abcam 15580) for 30 min. at room temperature, and Ki67+ nuclei were detected using Biocare Medical’s Mach 2 detection kit (MRCT523). Ki67 positive nuclei/mm2 were counted in each tumor and averaged within each group using an Olympus BX51 microscope fitted with an Olympus DP72 digital camera and CellSens Entry software (Olympus) to calculate tumor area.
Dv2004g1
Manufactured by Biocare Medical
The DV2004G1 is a laboratory equipment product designed for general laboratory use. It serves as a centrifuge, a device used to separate different components of a liquid mixture based on their relative densities. The DV2004G1 model is capable of performing this core function, but no further details about its intended use or specific features are provided.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dv2004g1
1
Quantifying Tumor Cell Proliferation
2
Lung Adenoma Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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5 μm lung sections from urethane, iloprost, and saline groups were deparaffinized, blocked with 0.3% hydrogen peroxide, and antigen retrieval performed in boiling Diva Decloaker (Biocare Medical, DV2004G1) under pressure for 5 minutes. Sections were blocked with Background Punisher (Biocare Medical BP974M) and 2.5% Normal Horse serum (Vector Labs S-2012-50). Sections were incubated in Ki67 primary Ab (1:2000 dilution, Abcam 15580) for 1 hour at room temperature, followed by anti-rabbit universal antibody for 30 minutes and ABC reagent for 30 minutes at room temperature (Vector Laboratories PK-7200). Ki67+ nuclei were detected using Betazoid DAB chromagen kit (Vector Laboratories BDB2004). Tumor area was measured and Ki67-positive nuclei/mm2 were counted in each tumor. Replicate blinded counts were conducted. The Ki67+ nuclei/mm2 tumor area for each tumor was averaged by group (Dwyer-Nield et al., 2017 (link)). H&E stains were done on 5 μm lung sections from urethane, iloprost, and saline groups and were used to confirm adenoma presence and structure.
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