How do you transfect S2 cells?

During transfection and selection keep cells in the same culture vessel. For general maintenance of cells, pass S2 cells when cell density is between 6 to 20 x 106 cells/ml and split at a 1:2 to 1:5 dilution. Note: S2 cells do not grow well when seeded at a density below 5 x 105 cells/ml.

How do you prepare cells for immunostaining?

Cell preparation for adherent cells

  1. Seed 1–1.5 x104 cells per well of a 4-chamber slide in 500 mL of culture medium. Incubate at 37°C at 5% CO2.
  2. 32–36 hours post cell seeding, remove the cell culture medium and rinse the cells 3 times using 500 µL of 1X PBS.

What are S2 cells used for?

S2 cells are often used for expression of heterologous proteins and can be used for large-scale production of proteins. Additionally, the cells can be easily transiently transfected with several plasmids at once to study protein interactions.

How do you fix a cell immunostaining?

Fixation. The cells may be fixed using one of two methods: Incubating the cells in 100% methanol (chilled at -20°C) at room temperature for 5 min. Using 4% paraformaldehyde in PBS pH 7.4 for 10 min at room temperature.

What is S2 library?

The S2 library defines a framework for decomposing the unit sphere into a hierarchy of cells. Each cell is a quadrilateral bounded by four geodesics.

What are Sf9 cells used for?

Sf9 cells, a clonal isolate of Spodoptera frugiperda Sf21 cells (IPLB-Sf21-AE), are commonly used in insect cell culture for recombinant protein production using baculovirus. They were originally established from ovarian tissue. They can be grown in the absence of serum, and can be cultured attached or in suspension.

What is immunostaining used for?

Immunostaining is used in cell biology to study differential protein expression, localization and distribution at the tissue, cellular, and subcellular level.

What does it mean to permeabilize a cell?

Permeabilization. The permeabilization step removes more cellular membrane lipids to allow large molecules like antibodies to get inside the cell.

What is S2 geometry?

S2 is a library for spherical geometry that aims to have the same robustness, flexibility, and performance as the very best planar geometry libraries.

What are S2 cells biology?

S2 cells are phagocytic cells that easily take up large amounts of DNA from the cell culture. They, thus, have a high cotransfection rate, allowing the coexpression of up to 12 different proteins.

What is the purpose of fixation during immunostaining?

What is the purpose of fixation during immunostaining? preserving cells prior to undergoing anitbody treatment.

How many S2 cells are there?

We now return to the question of how the S2 cells are numbered. Recall that there are 31 levels of subdivision, ranging from 0 to 30.