Method: - 293 GT cells
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Method: - 293 GT cells - average count of 1:2 and 1:4 dilutions was used to prepare cell titration (20,000, 5,000, 1,000, and 200) - cells seeded in morning, allowed to settle and adhere for 4 hours - CellTiter-Glo in white plate (Perkin Elmer Isoplate-96 TC 1450-516), CyQUANT in black plate (Costar 3603) - CTG was read twice, once after 2 min shake and 10 min RT equilibration and then again 30 min after first read - CyQUANT was read after 30 min at 37oC Notes: - CTG is the easier of the two. Requires 2 min orbital shaking which could be done in the EnVision. Very linear response from 0 to 20000 cells/well and very tight replicates too. - CyQUANT is a little more involved. Requires removal of media and longish incubation at 37oC (30-60min). Also, according to Invitrogen this incubation time is dependant on the cell type so it would have to be optimized specifically. Test of CellTiter-Glo and CyQUANT cell assay kits
description
Test of CellTiter-Glo and CyQUANT cell assay kits. Method: - 293 GT cells - average count of 1:2 and 1:4 dilutions was used to prepare cell titration (20,000, 5,000, 1,000, and 200) - cells seeded in morning, allowed to settle and adhere for 4 hours - PowerPoint PPT Presentation
Transcript of Method: - 293 GT cells
Method:
- 293 GT cells- average count of 1:2 and 1:4 dilutions was used to prepare cell titration (20,000, 5,000, 1,000, and 200)- cells seeded in morning, allowed to settle and adhere for 4 hours- CellTiter-Glo in white plate (Perkin Elmer Isoplate-96 TC 1450-516), CyQUANT in black plate (Costar 3603)- CTG was read twice, once after 2 min shake and 10 min RT equilibration and then again 30 min after first read- CyQUANT was read after 30 min at 37oC
Notes:
- CTG is the easier of the two. Requires 2 min orbital shaking which could be done in the EnVision. Very linear response from 0 to 20000 cells/well and very tight replicates too.- CyQUANT is a little more involved. Requires removal of media and longish incubation at 37oC (30-60min). Also, according to Invitrogen this incubation time is dependant on the cell type so it would have to be optimized specifically.
Test of CellTiter-Glo and CyQUANT cell assay kits
Read 1
y = 487.08x + 53928
R2 = 0.9996
0
2000000
4000000
6000000
8000000
10000000
12000000
0 5000 10000 15000 20000 25000
Cell count
CPS
Read 2
y = 487.75x + 54986
R2 = 0.9996
0
2000000
4000000
6000000
8000000
10000000
12000000
0 5000 10000 15000 20000 25000
Cell count
CPS
Read 3
y = 486.28x + 55326
R2 = 0.9996
0
2000000
4000000
6000000
8000000
10000000
12000000
0 5000 10000 15000 20000 25000
Cell count
CPS
Read 1
y = 465.12x + 69328
R2 = 0.9994
0
2000000
4000000
6000000
8000000
10000000
12000000
0 5000 10000 15000 20000 25000
Cell count
CPS
Read 2
y = 454.73x + 134833
R2 = 0.9997
-2000000
0
2000000
4000000
6000000
8000000
10000000
0 5000 10000 15000 20000 25000
Cell count
CPS
Read 3
y = 424.92x + 384962
R2 = 0.9999
-2000000
0
2000000
4000000
6000000
8000000
10000000
0 5000 10000 15000 20000 25000
Cell count
CPS
CellTiter-Glo
20,000 5,000 1,000
200 0
CellTiter-Glo
Read 1
y = 925.4x + 576328
R2 = 0.9999
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
0 1000 2000 3000 4000 5000 6000
Cell count
RFU
Read 2
y = 924.71x + 563018
R2 = 0.9997
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
0 1000 2000 3000 4000 5000 6000
Cell count
RFU
Read 3
y = 906.21x + 573763
R2 = 0.9997
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
0 1000 2000 3000 4000 5000 6000
Cell count
RFU
CyQUANT
20,000 5,000 1,000
200 0
CyQUANT
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