Laboratory methods

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Laboratory methods

Post by lrob » Sun Mar 06, 2011 1:44 pm

Hi, I have been reading a paper, and understand the aims and outcomes of the study, but don't understand the methods they use. Why do they use these techniques?
Here's a link: ... 7049.x/pdf

Here's the reference:
Maiso, P. et al. 2007. The insulin-like growth factor-I receptor inhibitor NVP-AEW541 provokes cell cycle arrest and apoptosis in multiple myeloma cells. British Journal of Haematology. 141, 470–482

Here's my (patchy) interpretation:
Cell proliferation, cell cycle and apoptosis assays- They are analysing the normal growth of the MM cells as a control

Microarray RNA analyses- To find out when the beginning of apoptosis is, and extract the total RNA at this point-but don’t know why

Immunoprecipitation and Western blotting- not sure what they’re trying to extract and analyse here.

Subcellular fractionation- Again, not too sure here.

Effect of interleukin-6, IGF-I and BMSC on NVP-AEW541-induced growth inhibition- This is to see what effect these chemicals have on NVP-AEW541

In vitro kinase assay- an in vitro test on the effects of NVP-AEW541 on the MM cells

MM xenografts- An in vivo test on a mouse

Any help on explaining the methodology of this paper would be very helpful- Thanks!

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Inland Taipan
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Post by JackBean » Mon Mar 07, 2011 8:07 am

you should read the results, there you should find, which experiment was used to explore which part.

blots: look e.g. on Fig. 3, there you find, what were they blotting...

fractionations - they were probably interested in which organelle are some of their proteins. But again look ito that article

Cis or trans? That's what matters.

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