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Revision as of 19:00, 1 July 2010 (view source) AndreasConstantinou (Talk | contribs) ← Older edit		Revision as of 15:38, 3 July 2010 (view source) JohanNordholm (Talk | contribs) m (→Hassan) Newer edit →
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	''Continued 30/6''		''Continued 30/6''
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		+	=Hassan=
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		+	''"It has become dangerously acceptable to conclude that proteins interact directly provided that (i) the interaction is biologically plausible; (ii) the proteins are co-expressed; and (iii) an interaction is demonstrated by glutathione-S-transferase (GST)-pulldowns and/or co-immunoprecipitations (co-IPs)"... "The most obvious caveat with co-IP data is that a positive result does not imply a direct interaction between two proteins – binding could be mediated by other partners."..."In addition, a widespread misconception is that co-IPs from extracts provide in vivo evidence for the existence of an interaction. This is not accurate, particularly when the experiment is carried out using overexpressed proteins in cell lines."...""
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		+	'' ""
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		+	"Protein interactions: is seeing believing? J. P. Mackay et al. 2007"

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Revision as of 15:38, 3 July 2010

Contents 1 Andreas 1.1 Expression of SOD and yCCS from pEX expression vector 1.1.1 Procedures 2 Hassan

Andreas

Expression of SOD and yCCS from pEX expression vector

Two cell cultures of DH5alpha cells respectively transformed with pEX vectors carrying superoxidase dismutase (SOD) and yeast copper chaperone (yCCS) were previously prepared and received from Nina and Johan.

Procedures

• An ON starter culture was set by inoculating 5 ml LB + 100 ug/ml Amp with 5 ul of each culture.
  • 37°C, 250 rpm.

Continued 30/6

Hassan

"It has become dangerously acceptable to conclude that proteins interact directly provided that (i) the interaction is biologically plausible; (ii) the proteins are co-expressed; and (iii) an interaction is demonstrated by glutathione-S-transferase (GST)-pulldowns and/or co-immunoprecipitations (co-IPs)"... "The most obvious caveat with co-IP data is that a positive result does not imply a direct interaction between two proteins – binding could be mediated by other partners."..."In addition, a widespread misconception is that co-IPs from extracts provide in vivo evidence for the existence of an interaction. This is not accurate, particularly when the experiment is carried out using overexpressed proteins in cell lines."...""

""

"Protein interactions: is seeing believing? J. P. Mackay et al. 2007"