Investigation of identity

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

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


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


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


Further tests on effects of VEGF

Diff concs

Effect of Anti-VEGF

Make Cltx conjugate

Cross linking exps and analysis



Cltx/VEGF/ anti VEGF treatment combos


Contingency/replicate week


Contingency/replicate week

Additional exps

Contingency/replicate week

Additional exps