Reagents and instruments for immunology, cell biology and molecular biology.
 
> Technical resources > Protocols > Cell culture > Culture and plate screening protocol for mammalian and insect cells

Culture and plate screening protocol for mammalian and insect cells

Culture and plate screening protocol for mammalian and insect cells

The plates represent a cross section of plates for different types of tests, cell growth and storage. We offer several formats: 96 and 24 wells to fit your needs.

 

Protocol for the screening of mammalian and insect cells - 96-well plates :
 
Material :
  • 96-well plate, 2 ml, square wells, round bottom, individually wrapped with lid, sterile : 931134
  • 96-well plate, 2 ml, square wells, pyramid bottom, individually wrapped with lid, sterile : 931133
 
Method :
 
1. Maintain cell stocks in appropriate growth medium. Divide cultures the day before transfection at an appropriate density to ensure log phase growth at the time of transfection.
2. Apply 500 ul / well of cell suspension. The optimal seeding density depends on the cell line, please use the recommended density for the cell line used.
3. Transfect cells according to the established transfection protocol. Establish the ratio of transfection reagent to DNA to nutrients from what is used for larger scale cultures.
4. Seal the plates with plastic covers or gaskets and transfer them to a shaker overnight at 800 rpm and 37°C.
5. Harvest cells at the established time for larger scale cultures. Pellet cells by centrifugation at 2500 g for 20 min at 4°C.
6. Reserve the culture medium or pellet according to the application and proceed with downstream processing.
 
Notes :
 
  • The most critical factor for cell viability is aeration. Optimal results will be obtained using shakers with orbital diameters of 3 mm. We do not recommend working in 96-well plates using shakers with standard diameters of 25 mm.
  • Thomson filtration plates are an excellent complementary product for downstream purification applications.

                 Filter plate 96 wells, 2 ml | 25µm Polypropylene: ref. 931919 

                 Suggested maximum centrifugation: 3000 g.

Protocol for the screening of mammalian and insect cells - 24-well plates :
 
Material :
  • 24-well plate, 10.4 ml, square wells, round bottom, individually wrapped with lid, sterile:  931568
  • 24-well plate, 10.8 ml, square wells, pyramid bottom, individually wrapped with lid, sterile :  931571
 
Method :
 
1. Maintain cell stocks in appropriate growth medium. Divide cultures the day before transfection at an appropriate density to ensure log phase growth at the time of transfection.
2. Apply 4-5 ml / well of cell suspension. The optimal seeding density depends on the cell line, please use the recommended density for the cell line used.
3. Transfect cells according to the established transfection protocol. Establish the ratio of transfection reagent to DNA to nutrients from what is used for larger scale cultures.
4. Seal the plates with plastic covers or gaskets and transfer them to a shaker overnight at 800 rpm and 37°C.
5. Harvest cells at the established time for larger scale cultures. Make a cell pellet by centrifugation at 2500 g for 20 min at 4 ° C.
6. Reserve the culture medium or pellet according to the application and proceed with downstream processing.
 
Notes :
 
  • The most critical factor for cell viability is aeration. Optimal results will be obtained using shakers with orbital diameters of 3 mm. We do not recommend working in 96-well plates using shakers with standard diameters of 25 mm.
  • Thomson filtration plates are an excellent complementary product for downstream purification applications.
                Filter plate 24 wells, 10.8 ml | 25µm Polypropylene: ref. 921550