How do you lyse cells in a 96-well plate?

The cells were treated with 90 μL per well of either 10% DMSO or compound solution and shaken gently at 37°C for 1 or 2 h. Following drug treatment, the cells were lysed in the wells.

How do you split cells in 96 well plate?

How do you remove media from a 96 well plate?

How do you Lyse adherent cells?

How do you seed cells evenly in 24 well plate?

It is a common issue due to vibration in incubators. Easiest method to overcome this issue – after adding the cell suspension to wells (minimal volume till cells get attached) gently swirl the plate to spread the cells evenly. Keep the plate on a flat surface for 5 minutes (bench top in the tissue culture room) .

How do you count and seed cells?

Take individual cell counts of all boxes, add them up and average them. Multiply the average with your dilution factor (in this case 10). This is the amount of cells in million per mL of your culture.

How do you get rid of bubbles in cell culture?

To get rid of the bubbles, try sparging through a CO2 permeable silicone membrane. The book “Cell and Tissue Engineering” written by D. Eibl is a valuable source to tackle this problem, too.

How do you remove bubbles from Elisa plate?

Sometimes air, resulting in bubbles, can be drawn into the pipette or dispensed into the wells. If this happens, bubbles can influence optical density values and results. To minimize or eliminate this problem, reverse pipetting is recommended for the addition of reagents to the ELISA plate.

How do you prepare a protein extraction buffer?

Protein Extraction Protocol Steps

Scrape the cells using cold plastic cell scraper. Collect the cells in microcentrifuge tubes. Agitate the contents in microcentrifuge tubes for 30 min at 4 °C. Centrifuge the tubes at 16,000 x g for 20 min at 4 °C.

How do you make a protein extraction buffer?

How do you find the surface area of a petri dish?

If, for instance, the actual lenght (i.e. diameter) of an stated 100 mm x 20 mm petri dish was 100 mm, it wouldn’t make sense that its surface area was 58 cm2. Rather, it should be π * R^2 / (3,14 * 50^2) = 78,53 cm^2.

How do you start a cell culture?

How is transfection done?

Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by electroporation, by cell squeezing, or by mixing a cationic lipid with the material to produce liposomes that fuse with the cell membrane and deposit their cargo inside.

How do you plate cells in a 96 well plate?

Cell doubling time is an important factor to be considered when adjusting cell density at the beginning of an experiment. Plate 200 µL of cell culture (i.e., 10,000–50,000 cells) into the wells of the sterile 96-well cell culture plate. Incubate the cells for 18 hours at 37°C. Stimulate the cells as desired.

How do you clean a hemocytometer?

How do you plate cells evenly?

How do you make a pipette without bubbles?

Focus on angles: To ensure you dispense all the liquid in your popette and avoid air bubbles, aspirate at a 90 degree angle and dispense at a 45 degree angle. Release pipettes slowly: After dispensing the liquid in your pipette, you shouldn’t release the plunger too quickly.

How do you make a western blot sample?

How do you lyse a cell?