The HK1 cells were seeded into 100-mm culture plates at a density of 4.0×105 cells/dish in 6 ml culture medium. The cells were treated with GnRH at a concentration of 10-9 M for 48 h. Untreated cells were used as a control. The cells were prepared for cell-cycle analysis using a CycleTEST PLUS DNA Reagent kit (BD BioSciences, San Jose, CA, USA). A pellet containing 5×105 cells was gently resuspended in 250 μl solution A, containing trypsin (BD Biosciences), followed by 200 μl solution B, containing trypsin inhibitor and RNase A (BD Biosciences), and incubated at room temperature for 10 min each. A total of 200 μl propidium iodide (PI) was added and the cell suspensions were incubated at 4°C in the dark for 10 min. Flow-cytometric analysis of the cellular DNA content was performed using Cell Quest Pro software version 6.0 (BD Biosciences) on a FACS Calibur flow cytometer (BD BioSciences) and the results were analysed using ModFit LT™ software version 4.0 (Verity Software House, Inc., Topsham, ME, USA).
Trypsin inhibitor
Manufactured by BD
Trypsin inhibitor is a type of enzyme inhibitor that specifically binds to and inactivates the enzyme trypsin. Trypsin is a protease enzyme involved in the digestion of proteins. The trypsin inhibitor helps regulate trypsin activity and is commonly used in laboratory settings to control proteolytic processes.
Automatically generated - may contain errors
Lab products found in correlation
100 μm nylon cell strainer, by BD (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
70 μm nylon cell strainer, by BD (2 mentions)
Soybean trypsin inhibitor, by Thermo Fisher Scientific (2 mentions)
Shandon cytospin, by Thermo Fisher Scientific (2 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cycletest plus dna reagent kit, by BD (1 mentions)
Rnase a, by BD (1 mentions)
Trypsin, by BD (1 mentions)
Cellquest pro software version 6, by BD (1 mentions)
Modfit lt software version 4, by Verity Software House (1 mentions)
3 protocols using trypsin inhibitor
1
GnRH-Induced Cell Cycle Analysis
2
Neonatal Mouse Epidermal Cell Fractionation
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The epidermis was collected from newborn mice and young pups up to 5.5 days of age as described above. For newborns, the basal and spinous keratinocytes-enriched fraction was obtained by incubating the epidermis in 0.05% trypsin-EDTA (Gibco, Grand Island, NY) at 37°C for 20 min in a small dish with the skin surface up followed by manually shaking the digested epidermis in cold PBS vigorously 20 times and filtering through a 70-μm nylon cell strainer (BD Biosciences, San Jose, NJ) into a tube containing cold soy bean trypsin inhibitor (Gibco) at 0.5 mg/ml in PBS. The granular keratinocytes-enriched fraction was obtained by incubating the residual epidermal sheet in 0.25% trypsin-EDTA (Gibco) at 37°C for 20 min in a small dish with the skin surface up followed by manually shaking the digested sheet in cold trypsin inhibitor solution vigorously 20 times and filtering through a 100-μm nylon cell strainer (BD Biosciences). Both cell fractions were then centrifuged at 800 rpm for 5 min at 4°C, resuspended in cold PBS, and spun onto slides at 400 rpm for 3 min in a Shandon Cytospin (Thermo Scientific, Rockford, IL) to make a cell monolayer. Slides were air dried for 10 min and stored at −20°C for up to a week.
3
Neonatal Mouse Epidermal Cell Fractionation
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