KLF2+/− mice (B6; 129S4‐Klf2tm1.1Hhn/J, Stock# 026926) were purchased from JAX Laboratory (Bar Harbor, ME). Mouse lung ECs were isolated as described previously.26 Briefly, lungs from 3 mice of each group were minced into pieces and digested with prewarmed type I collagenase (2 mg/mL) at 37°C for 45 minutes. The suspension was filtered through a 70‐μm disposable cell strainer and then centrifuged at 800 g at 4°C for 5 minutes. Precipitates were resuspended in cold PBS+BSA+Penicillin/Streptomycin. The cell suspension was then incubated with CD31 (#553370; BD Pharmingen, San Jose, CA)‐coated Dynabeads (Invitrogen, Carlsbad, CA) on a rotor at room temperature for 15 minutes, and then washed until the suspension became visibly clear. Cells with beads were resuspended in growth medium (DMEM+20%FBS+penicillin/streptomycin+heparin) and plated on 0.1% gelatin‐coated culture dishes. At 5 to 9 days after plating, cells approached confluence. A second sort of cells were performed with CD102‐coated (#553326; BD Pharmingen) Dynabeads sorting. Then, cells were cultured in growth medium and used for indicated experiments immediately.
Cd102
Manufactured by BD
Sourced in Denmark
The CD102 is a laboratory instrument designed for the analysis and processing of biological samples. It features a compact and robust design, enabling consistent and reliable performance. The core function of the CD102 is to provide accurate and reproducible results in a variety of laboratory applications.
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Lab products found in correlation
2 protocols using cd102
1
Isolation and Characterization of Mouse Lung Endothelial Cells
2
Isolation and Cultivation of Cardiac Endothelial Cells
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A fresh isolated CC strip was placed in sterile PBS and cut into small pieces together with collagenase II (400 mg ml−1, Worthington Biochemical Corporation, Lakewood, NJ, USA) for 60 min. at 37°C. The sampled tissue was centrifuged at 900 RPM with isolation of the pellet and transferred to a coated gelatin and collagen petri dish at 37°C. The culture media was Dulbecco's Modified Eagle Medium with Fungizone, penicillin streptomycin, new born fetal calf serum and HEPES. When the cells had reached confluence after 1–4 days, the cells were incubated with the endothelium specific CD102 [intracellular adhesion molecule 2 (ICAM2)] rat anti-mouse (BD Biosciences, Albertslund, Denmark) for 30 min followed by incubation with Dynabeads with sheep anti-rat IgG (Life Technologies, Oslo, Norway) for 30 min at 4°C. Trypsin was added to separate the cells from the petri dish and transferred to a dynamag 5TM (Life Technologies, Oslo, Norway) for a 2 min clean, the pellet was dissolved and left in a coated petri dish and further cultivation was performed in endothelial cell growth serum (Provitro AG, Berlin, Germany) until patch clamp experiments were performed.
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