Sumanpreet- general project plan
Investigation of identity of Cltx membrane binding partner in MDA MB-231 and U251-MG cancer cell lines
Investigate the effects of VEGF and ClTx on cancer cell proliferation
Investigate NRP1 as CXltx membrane-binding partner on cancer cell lines
General plan
| Week | Cell culture strand | Western blotting and Cltx binding partner strand |
| 1 |
Get cells growing Practice counting cells Trypan blue assay |
Getting started in the lab sheet BCA protein assay practice SDS PAGE practice with FCS or BSA |
| 2 |
Initial cell titre glo assay using 5000 cells/well in serum and in serum free media. |
Make cell lysates Calculate protein concentration Read up about total protein staining for WB standardisation |
| 3 |
May need to adjust cell number Effect of VEGF in +/- serum conditions |
WB for NRP1 Check expression in your cell lines Work out how much lysate (and cells) need for later exps |
| 4 |
Further tests on effects of VEGF Diff concs Effect of Anti-VEGF |
Make Cltx conjugate Cross linking exps and analysis Internalisation |
| 5 |
Cltx/VEGF/ anti VEGF treatment combos |
|
| 6 |
Contingency/replicate week |
|
| 7 |
Contingency/replicate week Additional exps |
Contingency/replicate week Additional exps |