Hi
I need to lyse THP1 monocytes and then use western blotting to quantify the levels of certain proteins in the lysate. I tried lysing the cells with RIPA buffer, but the high detergent concentration was incompatable with running a gel. I then tried Lamberts Break buffer, but the total protein concentration of my lysare was extremely low (i.e. the cells didnt lyse).
Does anyone have any suggestions ass to how I can successfully lyse THP1s in a buffer that is compatable with SDS PAGE??
Thank you in advance!!
0
1 reply to this topic
#2
GNANA
-
- Global Moderators
-
- 195 posts
Veteran
7
Neutral
Neutral
Posted 01 September 2011 - 06:26 AM
I dont think RIPA would interfere your gel run!! post your RIPA composition and also tell how it interferes in the running of lysate...
I would prefer being perfectionist rather than a passionist in Research.
I always had an alternate hypothesis....
I always had an alternate hypothesis....
Also tagged with one or more of these keywords: Cell lysis, THP1
Protocols and Techniques Forums →
Protein and Proteomics →
Low pH Cell Lysis BufferStarted by Guest_ahitron_* , 02 Jan 2013  cell lysis, ph, SDS-PAGE
|
|
|
||
Protocols and Techniques Forums →
Tissue and Cell Culture →
IGF1 stimulation of THP1 cellsStarted by Guest_dude3005_* , 28 Sep 2012 IGF1, Cell culture, THP1, doping
|
|
|
||
Protocols and Techniques Forums →
Protein and Proteomics →
PopCulture Reagent and LC/MS/MS?Started by Guest_miche_* , 20 May 2012 LC MS, cell lysis
|
|
|
||
Protocols and Techniques Forums →
Cell Biology →
The connection between LPS and PMA stimulation for maturation of ThP1 cellsStarted by Guest_nat2012_* , 08 May 2012 ThP1, LPS, IL-18, PMA
|
|
|
||
Protocols and Techniques Forums →
Tissue and Cell Culture →
THP1 monocytes cultured forms "vacuols-like" compartmentsStarted by Guest_Exvitae_* , 01 Mar 2012 THP1, vacuols
|
|
|
